Using the Nikon TE2000 Inverted Microscope

Size: px
Start display at page:

Download "Using the Nikon TE2000 Inverted Microscope"

Transcription

1 Wellcome Trust Centre for Human Genetics Molecular Cytogenetics and Microscopy Core Using the Nikon TE2000 Inverted Microscope Fluorescence image acquisition using Scanalytic s IPLab software and the B&W Hamamatsu Orca C camera INSTRUCTIONS PLEASE HANDLE CAREFULLY! Handle the microscope gently, taking care to avoid sharp impacts. 1) TO SWITCH-ON THE MICROSCOPE AND LOOK AT YOUR SAMPLE: Bright-field Microscopy: - Turn on the transmitted light by turning on the switch on the Nikon power supply box on the bench to the left side of the microscope. - Press lamp on/off button at the base of the microscope on the left. - Adjust the light intensity by rotating the control dial situated on the left side of the microscope. - Check that the D (Diffuser) and HA (Heat Absorber) filters at the top left of the microscope are pushed in. Select the optional neutral density ND filters as necessary to dim the light intensity. Push in the optional [top right] neutral density ND16 and ND2 filters to dim the light intensity - push both in for maximum light attenuation, which often the optimal setting. - The eyepiece turret dial should be set to O (open), when set to C it is closed. - The "port/optical path" dial on the right side of the base should be set to port 1. - The "reticule in/out" lever at front of the base to the right should be turned clockwise and set to a 2 o clock position. - The fluorescence filter wheel located underneath the objective wheel should be on the empty position indicated by the double arrow symbol ( ). - Place the culture vessel or glass slide on the stage. 1

2 - Move a low power objective (4x or 10x) into the light path. Set the optical zoom knob [left side] to 1.0x or 1.5x as required. - Have the phase ring set to A for stained cells or Ph1 for phase when using the 4x, 10x and 20x objectives or Ph2 for phase when using the 40x and 60x objectives. DIC H is for use with the 60x oil objective only. - A hi-resolution 60x oil objective is also available if required [for glass slides]. If you need to use this please ask a member of core staff to attach it for you - Glass microscope slides are normally viewed coverslip-side down. Focus with the coarse focusing wheel, revolve to the 40x or 60x objective as required. - Please take extra care to avoid sharp impacts when switching between objectives. You may need to adjust the height of the objective wheel and refocus. - Focus on the specimen. Adjust the condenser to the correct height for Koehler illumination. Ask core staff for help with this if necessary. - The 20x, 40x and 60x objectives have correction collars to compensate for differences in vessel wall thickness. The collars can be adjusted by rotating them, and, used in conjunction with the normal focus, an optimal image can be obtained. Please ask Core staff for help with adjusting correction collars if necessary. The 60x oil objective with DIC [Differential Interference Contrast] Microscopy: - Ask a member of core staff to attach the 60x oil objective and set up the transmitted light DIC optics [if required]. - Have the phase ring set to DIC H. - Slide the polariser filter [located above the condenser turret] to the left. - Push in the black T-A analyser slider [located under the objectives & filter wheel, to the left of the microscope base]. - Place microscope slide cover-slip side down. Move a low power objective (4x or 10x) into the light path. When you have found an area of interest carefully revolve to the 60x oil objective. - Place a drop of oil on the objective. Make sure you use the correct oil. Only Cargille oil should be used with this objective. - For correct DIC you should have set the condenser height correctly for Koehler illumination. You then rotate the polariser, analyser and slide in/out the DIC prism [under the 60x objective] to give the best contrast enhanced image. 2

3 Epi-Fluorescence Microscopy: - Note the last switch-off time in the notebook beside the microscope. The bulb must be left for at least 30 minutes to cool down before it is switched on again. - Turn the power switch on the front panel of the mercury lamp power supply box (on the bench on the right side of the microscope) and make sure the "power" monitor light (green) is on. Always switch the mercury lamp on and off with the PC, LCD display, microscope and camera switched OFF. - Press the ignition button for 2-3 seconds to light the lamp. The "lamp ready" light (orange) will come on briefly. Leave to warm up for few minutes, the orange light should then be on permanently. Leave the mercury lamp on for at least 30 minutes before switching it off. - Unless you also capturing phase contrast or transmission images as well, the height and settings of the condenser above the stage are irrelevant for epifluorescence imaging. Fluorsescence Filter Sets Optical filters are required in the microscope to ensure appropriate excitation of the fluorophore. These dichroic filter sets are moved into the fluorescent light path by rotating the fluorescence filter wheel, which is located underneath the objective wheel [nosepiece], on the right side of the microscope base. The following filter sets are available: The following filter sets are available: o DAPI/uV Hoechst - Blue emission colour [violet excitation] o FITC/GFP GFP, Alexa Green emission colour [blue excitation] o TRITC/Red Rhodamine, Alexa Red emission colour [green excitation] o CY5 CY5, Alexa Far Red emission colour [red excitation] o Transmitted light only - Fluorescence off [Phase Contrast] Note: the excitation light colour is seen at the specimen and the emission light colour is seen down the microscope via the eye-pieces. - During fluorescence microscopy, turn off the bright-field halogen light by using either the switch on the left side of the microscope main body, or by rotating the transmitted light intensity knob anti-clockwise to zero. - Rotate the fluorescence filter wheel to select the desired excitation filter set [DAPI, FITC, TRITC or CY5]. - There is a fluorescence light shutter underneath the filter wheel (at the right hand side) that needs to be opened. Set to the O position for the fluorescence light to pass to the objectives. In the C position the shutter is closed. The shutter should be kept closed [C] during transmitted light microscopy. 3

4 - Adjust the fluorescence light brightness by inserting the ND filters (ND4 and ND8) into the optical path. These are located near the rear UV lamp housing. 2) TO START THE CAMERA AND CAPTURE IMAGES: - Switch on the Hamamatsu camera by pressing the switch on the camera power supply box next the 24 computer monitor. Turn on the computer (PC on the floor under the bench) and its 24 LCD monitor (button top right edge). - If you are a Well user log on to the system and network with your usual username and password. Alternatively, log on directly to the PC as User with the password cyto02. - Double-click on the IPLab 37 icon. - Click on Camera, and then Select Camera and from the scroll down menu choose Hamamatsu 1394 ORCA Once you have found an image down the microscope that you wish to take a picture of, move the port/optical path dial (on the right side of the base of the microscope) to port 5 [all light to the Hamamatsu camera]. - Click again on Camera, and then select Acquire Focus. A live video image will appear on the screen. This configuration is intended to allow you to get the image in focus while viewing it on the computer monitor. - An Acquire Preview dialog box appears. You can improve the live image by altering the camera image acquisition parameters 1. - Adjust the camera exposure time as required. The time is listed in milliseconds. - You may adjust the image contrast by using the white and black point slider bars (White Point: the pixel value at which the image will appear white; Black Point: the image value at which the image will appear black). - By checking the Show Saturated Pixels box you will be able to see pixels that are saturated. The pixels in blue are equal to or below the saturated black point and the pixels in red are equal to or above the set white point [i.e. overexposed]. Remember to uncheck the box before taking a picture! - The Pause button will stop the preview without closing the window. - To take a picture click on Snapshot. 1 The Hamamatsu camera captures 12-bit B&W (4,096 grey levels) images by default, with a maximum resolution of 1344 x 1025 pixels [with binning set to 1x1]. This can be reduced to 672 x 512 pixels with binning set to 2x2 where every four pixels of the CCD are combined, doubling camera sensitivity and halving resolution. Camera binning is adjusted in the Acquire Preview window, under the Size tab. 4

5 3) TO CREATE AND SAVE MULTICOLOUR IMAGES: - Capture separate grey scale images for phase or DIC and for each different fluorophore [e.g. DAPI, FITC] - Click on main menu Ext, and then select Create Multiprobe Image. - Select a window, image file name and choose probe name and colour. - Select next window and repeat. - Merge images by clicking OK. - Click on Ext, and then select Enhance Multiprobe Image to adjust the contrast and positioning of images that have been merged. Min/Max: these controls allow you to manipulate the normalisation, or contrast settings, for each channel. Pixel Shift: these controls move the image of the selected channel in case the images initially produced were not perfectly registered. Blending: this affects to what degree overlapping colour channels are mixed together to create an intermediate colour (i.e. at 100% the channel with the highest intensity value will show through). - Clicking the OK button makes changes according to the dialog settings and then exit the dialog box. - Save images in your own folder in User Images in drive C: / - The Image Information button [ i ], built into the image window, displays a window of information about the image. You can go to File and then select Edit Image Info and type in additional notes (i.e. objective you used, etc.). - When any image is saved as an IPL file, you can return to the file at any time to view or edit it. IPLabs can only open images saved in this ipl format, it can t open images it has exported to TIF or jpg files. To be able to open your images with Photoshop you must create an RGB image by clicking on Ext, select Extract Multiprobe Data and then save your images (either as single channels or 24-bit colour) in a more common format - like TIFF or jpg. Alternatively, you must export 2 your images by clicking on Math, then select Convert to 8/24 bit and then Save as. You can check that the exported image looks OK using Photoshop Elements 7 on the PC. 2 Hamamatsu camera images are captured at 12-bit black and white (4,096 grey levels). Use Math, Convert to 8/24 bit to convert these 12-bit images to 8-bit [256 grey levels] B&W, if you want to view them correctly in PhotoShop or in PowerPoint. We can only distinguish up to 191 different grey levels in a B&W image. 5

6 4) TO FINISH ( and go home): - Close IPLab 3.7 before switching off the camera. In order to close IPLab 3.7 you must click the OK button in the Acquire Preview window before selecting Exit. - If using the 60x oil objective please remember to wipe off the oil and inform a member of core staff when you have finished, so the objective can be removed. Microscope oil objectives should only be cleaned with the lens tissue provided. - If used, press the halogen transmission lamp on/off button at the base of the microscope on the left and turn off the Hamamatsu camera power supply box. - Switch off the computer and its LCD monitor. - Before turning off the fluorescence mercury lamp allow the mercury bulb to run for a minimum of 30 minutes and check in the booking sheet on the door that nobody else will be using the microscope in the next few hours. - Switch off the mercury lamp power supply box and note the time off and total hours used in the notebook. If you have any microscope/imaging problems or queries, just ask a member of the Microscopy Core staff for help. There is additional help and advice on our Core web-pages: WTCHG Fluorescence image acquisition using Scanalytic s IPLab software and the B&W Hamamatsu Orca C camera. This version updated: November

7 TROUBLESHOOTING If you do not see any fluorescence, check the following: Is there a shadow in the image with fluorescence illumination? If the shadow doesn t move with the specimen, an incorrectly set filter slider or other component may be obscuring the light path. Alternatively, the mercury lamp may have a problem ask a core member for help. Is the mercury bulb turned on? Check the orange and green light on the power box. Can you see coloured light passing through the objective? If not, check that the shutter underneath the filter wheel is open. Is the filter block in place? Are you using the correct filter block? Rotate the filter wheel and see if any light of the right colour appears. Have you checked the dial below the eyepiece? It should be set to O. Is the objective correctly positioned? The objective wheel should click into position. Is the port/optical path in the correct position? 1 is for observation down the microscope, 5 is for camera use only! Is the light too faint or too bright? Remove/Add the neutral density filters. My exported images are all black in Photoshop and PowerPoint The Hamamatsu camera captures in 12-bit [4,096 grey levels]. Photoshop can only view 8-bit [256 grey level] B&W images correctly. Core staff can advise on image bit depth conversion. FOR ANY QUERIES OR DOUBTS, JUST ASK A MEMBER OF THE CORE STAFF 7

Nikon E800 Operating Instructions.

Nikon E800 Operating Instructions. Nikon E800 Operating Instructions. You can request electronic copies of this manual by contacting lshats@jhsph.edu Copies are also available on the JHU MMI Department web site. Please send your comments

More information

Nikon E800 Microscope. Operating Instructions

Nikon E800 Microscope. Operating Instructions Nikon E800 Microscope Operating Instructions B Watson 12/2005 Table of contents: 1. The Nikon E800 Microscope 2. Turning the system ON and OFF 3. Selecting the light path 4. Operating in transmitted light

More information

Zeiss Axio Imager.A1 manual

Zeiss Axio Imager.A1 manual Zeiss Axio Imager.A1 manual Power-up protocol 1. Mercury lamp 2. Power strip on shelf 3. Computer The Mercury lamp should always be first-on and last-off. This prevents any electrical surges caused by

More information

Widefield-NikonEclipseTE200-ORCA Nikon Eclipse TE200 Inverted Microscope with Hamamatsu 1394 Orca-ER Cooled CCD Camera and Micromanager Software

Widefield-NikonEclipseTE200-ORCA Nikon Eclipse TE200 Inverted Microscope with Hamamatsu 1394 Orca-ER Cooled CCD Camera and Micromanager Software Widefield-NikonEclipseTE200-ORCA Nikon Eclipse TE200 Inverted Microscope with Hamamatsu 1394 Orca-ER Cooled CCD Camera and Micromanager Software September 2007 Check website for most current User Guide

More information

Nikon E800 Operating Instructions.

Nikon E800 Operating Instructions. Nikon E800 Operating Instructions. You can request electronic copies of this manual by contacting imaging@fhcrc.org. Copies are also available on the Scientific Imaging web site. Please send your comments

More information

Guide to Confocal 5. Starting session

Guide to Confocal 5. Starting session Guide to Confocal 5 Remember that when booking and before starting session you can check for any problems at https://www.bris.ac.uk/biochemistry/uobonly/cif/index.html Starting session Switch on microscope

More information

CONFOCAL MICROSCOPE (Zeiss LSM 510 META v4.2)

CONFOCAL MICROSCOPE (Zeiss LSM 510 META v4.2) Wellcome Trust Centre for Human Genetics Molecular Cytogenetics and Microscopy Core CONFOCAL MICROSCOPE (Zeiss LSM 510 META v4.2) 1) STARTING THE SYSTEM Abridged INSTRUCTIONS Switch on the mercury bulb

More information

Contents STARTUP MICROSCOPE CONTROLS CAMERA CONTROLS SOFTWARE CONTROLS EXPOSURE AND CONTRAST MONOCHROME IMAGE HANDLING

Contents STARTUP MICROSCOPE CONTROLS CAMERA CONTROLS SOFTWARE CONTROLS EXPOSURE AND CONTRAST MONOCHROME IMAGE HANDLING Operations Guide Contents STARTUP MICROSCOPE CONTROLS CAMERA CONTROLS SOFTWARE CONTROLS EXPOSURE AND CONTRAST MONOCHROME IMAGE HANDLING Nikon Eclipse 90i Operations Guide STARTUP Startup Powering Up Fluorescence

More information

Things to check before start-up.

Things to check before start-up. Byeong Cha Page 1 11/24/2009 Manual for Leica SP2 Confocal Microscope Enter you name, the date, the time, and the account number in the user log book. Things to check before start-up. Make sure that your

More information

INSTRUCTIONS FOR COURSE WORK 4 (AxioVert) Instructor: Anne Vaahtokari (MIU) 1. Purpose of the work

INSTRUCTIONS FOR COURSE WORK 4 (AxioVert) Instructor: Anne Vaahtokari (MIU) 1. Purpose of the work INSTRUCTIONS FOR COURSE WORK 4 (AxioVert) Instructor: Anne Vaahtokari (MIU) 1. Purpose of the work In this work, you will get familiar with an inverted epifluorescence microscope. Also, you will learn

More information

EPIFLUORESCENCE &/OR BRIGHTFIELD MICROSCOPY

EPIFLUORESCENCE &/OR BRIGHTFIELD MICROSCOPY EPIFLUORESCENCE &/OR BRIGHTFIELD MICROSCOPY TURN ON THE FOLLOWING EQUIPMENT The fluorescent light (if needed) The power strip for the microscope and accessories The CoolSNAP HQ camera on the right (Turn

More information

Nikon Eclipse Ti A1-A Confocal Operating Manual. Start-up. Microscope

Nikon Eclipse Ti A1-A Confocal Operating Manual. Start-up. Microscope Nikon Eclipse Ti A1-A Confocal Operating Manual Start-up 1. Turn on Excite Fluorescent light power supply- metal halide. a. Cool down as for mercury bulb b. Wheel closed liquid light guide 2. Turn on power

More information

Nikon SIM-E & A1-R System

Nikon SIM-E & A1-R System Nikon SIM-E & A1-R System USER GUIDE LSU Health Sciences Center Shreveport Research Core Facility June 01 2017 Chaowei Shang 1 Table of Content 1. Start Up the System... Page 3 Hardware and microscope

More information

Instructions for Making On-Line Reservations for Microscopes in NB11-204

Instructions for Making On-Line Reservations for Microscopes in NB11-204 Instructions for Making On-Line Reservations for Microscopes in NB11-204 1. Log into Mail using Mail.swmed.edu 2. Log in using your university id and password. 3. Click the Calendar Tab at the top right

More information

Zeiss Deconvolution Microscope: A Quick Guide

Zeiss Deconvolution Microscope: A Quick Guide Zeiss Deconvolution Microscope: A Quick Guide Start-up Uncover microscope. Do not put dust cover on the floor. Plug in both cameras. The default camera is the AxioCam HRm (monochrome camera) for fluorescence

More information

Widefield 1. Switching on

Widefield 1. Switching on Widefield 1 Switching on 1. Ignite DG5 lamp - must be switched on first (if previous user has switched off, wait 30 min before igniting) 2. Wait 5s and then turn on the main DG5 controller switch. 3. DG5

More information

Cell Biology and Bioimaging Core

Cell Biology and Bioimaging Core Cell Biology and Bioimaging Core Leica TCS SP5 Operating Instructions Starting up the instrument 1. First, log in the log book located on the confocal desk. Include your name, your lab s PI, an account

More information

Motorized Axio Observer Start-up instructions

Motorized Axio Observer Start-up instructions Start-up instructions 1. If using fluorescence turn on Fluorescent light source. TL light Source (Hal 100) 2. Turn on microscope using switch on lower left side of the microscope. 3. If imaging, turn on

More information

Quick Start Guide. Leica SP5 X

Quick Start Guide. Leica SP5 X Quick Start Guide Leica SP5 X Please note: Some of the information in this guide was taken from Leica Microsystems Leica TCS SP5 LAS AF Guide for New Users. This work is licensed under the Creative Commons

More information

Characterization Microscope Nikon LV150

Characterization Microscope Nikon LV150 Characterization Microscope Nikon LV150 Figure 1: Microscope Nikon LV150 Introduction This upright optical microscope is designed for investigating up to 150 mm (6 inch) semiconductor wafers but can also

More information

User manual for Nikon Elements software

User manual for Nikon Elements software User manual for Nikon Elements software Equipment: Nikon TE300 Eclipse microscope ANDOR Neo/Zyla B&W camera (default) DS Fi2 color camera Sign in on the sign in sheet; please use both your given name and

More information

SHORT INSTRUCTIONS FOR OPERATING LSM1/2 (Zeiss LSM510) AT CIAN Version 1.4, September 2014

SHORT INSTRUCTIONS FOR OPERATING LSM1/2 (Zeiss LSM510) AT CIAN Version 1.4, September 2014 CIAN LSM1 or LSM2 short instructions, version 1.4, September 2014 page 1 of 6 SHORT INSTRUCTIONS FOR OPERATING LSM1/2 (Zeiss LSM510) AT CIAN Version 1.4, September 2014 Before starting To work with LSM1

More information

Nikon C1si Spectral Laser Scanning Confocal Microscope. User Guide

Nikon C1si Spectral Laser Scanning Confocal Microscope. User Guide Nikon C1si Spectral Laser Scanning Confocal Microscope User Guide Contents: C1Si Turn-On/ShutDown Procedures... 2 Overview... 4 Setup for epi-illumination to view through the eyepieces:... 5 Setup for

More information

Nikon Ti-E Microscope Manual. Rightmire Hall Ohio State University. Director: Tony Brown Rightmire

Nikon Ti-E Microscope Manual. Rightmire Hall Ohio State University. Director: Tony Brown Rightmire Nikon Ti-E Microscope Manual Rightmire Hall Ohio State University Director: Tony Brown Rightmire 060 292-1205 brown.2302@osu.edu Facility Manager: Paula Monsma Rightmire 062 293-0939 292-1367 monsma.1@osu.edu

More information

Leica SPEII confocal microscope. Short Manual

Leica SPEII confocal microscope. Short Manual Leica SPEII confocal microscope Short Manual Switching ON sequence: 1. Turn on the Workstation under the bench (top, far right). 2. Turn on the Supply Unit - Laser box (big green switch first and then

More information

Nikon TE300 Eclipse Wide-Field Microscope

Nikon TE300 Eclipse Wide-Field Microscope Nikon TE300 Eclipse Wide-Field Microscope User Guide LSU Health Science Center-Shreveport Research Core Facility 1 User manual for Nikon Elements software Equipment: Nikon TE300 Eclipse microscope Photometrics

More information

START-UP PROCEDURE 1 THE MICROSCOPE STAND 3 OBJECTIVES 5 STARTING WITH LAS (SOFTWARE) AND SETTING UP THE MICROSCOPE STAND 7

START-UP PROCEDURE 1 THE MICROSCOPE STAND 3 OBJECTIVES 5 STARTING WITH LAS (SOFTWARE) AND SETTING UP THE MICROSCOPE STAND 7 Leica DMI AF6000LX Table of contents START-UP PROCEDURE 1 THE MICROSCOPE STAND 3 OBJECTIVES 5 STARTING WITH LAS (SOFTWARE) AND SETTING UP THE MICROSCOPE STAND 7 ACQUIRE MODULE 6 SETTING THE LIGHTPATH 6

More information

ZEISS LSM510META confocal manual

ZEISS LSM510META confocal manual ZEISS LSM510META confocal manual Switching on the system 1) Switch on the Remote Control button located on the table to the right of the microscope. This is the main switch for the whole system including

More information

Olympus IX71 Microscope and DP71 Camera Instructions

Olympus IX71 Microscope and DP71 Camera Instructions Olympus IX71 Microscope and DP71 Camera Instructions Microscopy in Medicine (MiM) Core Emory University Department of Medicine 1 Olympus IX71 Image Capture Procedure 2 3 1. STARTING-UP PROCEDURE: Remove

More information

Zeiss Axiovert 135 Fluorescence Microscope Quick Guide / Operations Manual (v. 1.0 February 09)

Zeiss Axiovert 135 Fluorescence Microscope Quick Guide / Operations Manual (v. 1.0 February 09) University of Chicago Integrated Light Microscopy Core Dr. Vytas Bindokas, Director http://digital.bsd.uchicago.edu By: Christine Labno, Assistant Director Room: AB-129 Phone: 4-9040 Zeiss Axiovert 135

More information

Leica SP8 TCS Users Manual

Leica SP8 TCS Users Manual Version : 07/08/0 Leica SP8 TCS Users Manual Start up:. Turn the PC Microscope, Scanner Power, Laser Power, and the Laser Emission key to on (bottom right of desk).. Turn on the fluorescent lamp (top left

More information

Olympus Fluoview 1000S Spectral Confocal Microscope Introduction to the NRI-MCDB Microscopy Facility Spectral Confocal Microscope

Olympus Fluoview 1000S Spectral Confocal Microscope Introduction to the NRI-MCDB Microscopy Facility Spectral Confocal Microscope Olympus Fluoview 1000S Spectral Confocal Microscope Introduction to the NRI-MCDB Microscopy Facility Spectral Confocal Microscope Improved Optics More Lasers 405 diode 440 diode 488 Argon 515 Argon 559

More information

Zeiss LSM 880 Protocol

Zeiss LSM 880 Protocol Zeiss LSM 880 Protocol 1) System Startup Please note put sign-up policy. You must inform the facility at least 24 hours beforehand if you can t come; otherwise, you will receive a charge for unused time.

More information

Zeiss Axioskop II. The AIF's "routine" light microscope. (Installed 8/24/04)AxioCam installed July 11th 2005

Zeiss Axioskop II. The AIF's routine light microscope. (Installed 8/24/04)AxioCam installed July 11th 2005 Zeiss Axioskop II The AIF's "routine" light microscope. (Installed 8/24/04)AxioCam installed July 11th 2005 Featuring: Phase Contrast Darkfield DIC/Nomarski Brightfield Fluorescent filters for Dapi, FITC,Rhodamine

More information

Operating Instructions for Zeiss LSM 510

Operating Instructions for Zeiss LSM 510 Operating Instructions for Zeiss LSM 510 Location: GNL 6.312q (BSL3) Questions? Contact: Maxim Ivannikov, maivanni@utmb.edu 1 Attend A Complementary Training Before Using The Microscope All future users

More information

TRAINING MANUAL. Olympus FV1000

TRAINING MANUAL. Olympus FV1000 TRAINING MANUAL Olympus FV1000 September 2014 TABLE OF CONTENTS A. Start-Up Procedure... 1 B. Visual Observation under the Microscope... 1 C. Image Acquisition... 4 A brief Overview of the Settings...

More information

personal DELTAVISION (pdv)

personal DELTAVISION (pdv) GUIDELINES AND HINTS Version 1.3 (March 2015) personal DELTAVISION (pdv) Epifluorescence microscope from Applied Precision Inc.: The microscope can be found in room 1.320. For details see the architectural

More information

Operation Guide for the Leica SP2 Confocal Microscope Bio-Imaging Facility Hunter College October 2009

Operation Guide for the Leica SP2 Confocal Microscope Bio-Imaging Facility Hunter College October 2009 Operation Guide for the Leica SP2 Confocal Microscope Bio-Imaging Facility Hunter College October 2009 Introduction of Fluoresence Confocal Microscopy The first confocal microscope was invented by Princeton

More information

Brightfield Microscopy and Image Acquisition on Spotcam1. by Ryan Taylor/Nancy Kleene Last modified 10/02/05 by Birgit Ehmer

Brightfield Microscopy and Image Acquisition on Spotcam1. by Ryan Taylor/Nancy Kleene Last modified 10/02/05 by Birgit Ehmer Brightfield Microscopy and Image Acquisition on Spotcam1 by Ryan Taylor/Nancy Kleene Last modified 10/02/05 by Birgit Ehmer Log onto the computer. Enter your username and password to log onto the server.

More information

Leica SP8 TCS Users Manual

Leica SP8 TCS Users Manual Leica SP8 TCS Users Manual Follow the procedure for start up and log on as posted in the lab. Please log on with your account only and do not share your password with anyone. We track and confirm usage

More information

Zeiss AxioImager.Z2 Brightfield Protocol

Zeiss AxioImager.Z2 Brightfield Protocol Zeiss AxioImager.Z2 Brightfield Protocol 1) System Startup Please note put sign-up policy. You must inform the facility at least 24 hours beforehand if you can t come; otherwise, you will receive a charge

More information

BX-61: Brightfield Instruction /Continue to scroll for Fluorescent Instuctions

BX-61: Brightfield Instruction /Continue to scroll for Fluorescent Instuctions BX-61: Brightfield Instruction /Continue to scroll for Fluorescent Instuctions Starting up: Schematic of Olympus BX-61. 1. Turn on Olympus microscope power box (left of microscope) with toggle switch on

More information

Nikon AZ100. Laser Scanning Macro Confocal Microscope. Jordan Briscoe Adam Fries Kyle Marchuk Kaitlin Corbin. May 2017.

Nikon AZ100. Laser Scanning Macro Confocal Microscope. Jordan Briscoe Adam Fries Kyle Marchuk Kaitlin Corbin. May 2017. Nikon AZ100 Laser Scanning Macro Confocal Microscope Jordan Briscoe Adam Fries Kyle Marchuk Kaitlin Corbin May 2017 Contents 1 Introduction 2 2 Hardware - Startup 2 3 Software/Operation 4 3.1 Multidimensional

More information

Diskovery Spinning Disk Guide

Diskovery Spinning Disk Guide Diskovery Spinning Disk Guide qbi.microscopy@uq.edu.au Getting started The microscope and its peripherals (Fig. 1a) should always be turned on, but if they are not, turn them on in the following way: 1.

More information

Operating Checklist for using the Laser Scanning Confocal Microscope. Leica TCS SP5.

Operating Checklist for using the Laser Scanning Confocal Microscope. Leica TCS SP5. Smith College August 2010 Operating Checklist for using the Laser Scanning Confocal Microscope Leica TCS SP5. CONTENT, page no. Startup, 1 Initial set-up, 1 Software, 2 Microscope Specimen observation

More information

AxioVision 4.5 Brightfield Image Capture Procedure

AxioVision 4.5 Brightfield Image Capture Procedure AxioVision 4.5 Brightfield Image Capture Procedure 1. STARTING-UP PROCEDURE: Remove blue dust cover and place on shelf under microscope. Turn on the halogen lamp by pushing the switch at the back right

More information

LSM 710 Confocal Microscope Standard Operation Protocol

LSM 710 Confocal Microscope Standard Operation Protocol LSM 710 Confocal Microscope Standard Operation Protocol Basic Operation Turning on the system 1. Switch on Main power switch 2. Switch on System / PC power button 3. Switch on Components power button 4.

More information

Practical work no. 3: Confocal Live Cell Microscopy

Practical work no. 3: Confocal Live Cell Microscopy Practical work no. 3: Confocal Live Cell Microscopy Course Instructor: Mikko Liljeström (MIU) 1 Background Confocal microscopy: The main idea behind confocality is that it suppresses the signal outside

More information

CAPTURING IMAGES ON THE HIGH-MAGNIFICATION MICROSCOPE

CAPTURING IMAGES ON THE HIGH-MAGNIFICATION MICROSCOPE University of Virginia ITC Academic Computing Health Sciences CAPTURING IMAGES ON THE HIGH-MAGNIFICATION MICROSCOPE Introduction The Olympus BH-2 microscope in ACHS s microscope lab has objectives from

More information

Simplified Instructions: Olympus Widefield Microscope S1230

Simplified Instructions: Olympus Widefield Microscope S1230 Contents General Microscope Operation Simple Image Capture Multi-Wavelength Capture Z-Series Timelapse Combining Capture Modes Synopsis of Other Functions Pages 2-23 24-40 41-47 48-56 57-59 60-68 69-83

More information

Leica TCS SL Confocal Training. Neuroscience Imaging Core Staff. Core Director. Facility Manager

Leica TCS SL Confocal Training. Neuroscience Imaging Core Staff. Core Director. Facility Manager Leica TCS SL Confocal Training Neuroscience Imaging Facility The Ohio State University Rightmire Hall 614-292-1367 Staff Core Director Anthony Brown, Ph. D. 060 Rightmire Hall 614-292-1205 brown.2302@osu.edu

More information

LSM 510 Training Notes

LSM 510 Training Notes LSM 510 Training Notes Turning on the system Turn on the arc lamp, found on the bench top left of the microscope. This supplies light for epifluorescence for viewing your samples through the microscope.

More information

Olympus xcellence Software - basic user guide

Olympus xcellence Software - basic user guide Olympus xcellence Software - basic user guide This is a basic overview of setting up time lapse experiments using Olympus's xcellence software on BIU's IX81 inverted phase contrast system - the software

More information

Microscope Confocal LSM510 META

Microscope Confocal LSM510 META Microscope Confocal LSM510 META Welcome to the Zeiss LSM 510 Meta Confocal tutorial. Before using the LSM 510 META, Log off any other computer that is open with your personal login. You will need to put

More information

Zeiss LSM 780 Protocol

Zeiss LSM 780 Protocol Zeiss LSM 780 Protocol 1) System Startup F Please note the sign-up policy. You must inform the facility at least 24 hours beforehand if you can t come; otherwise, you will receive a charge for unused time.

More information

MAKE SURE YOUR SLIDES ARE CLEAN (TOP & BOTTOM) BEFORE LOADING DO NOT LOAD SLIDES DURING SOFTWARE INITIALIZATION

MAKE SURE YOUR SLIDES ARE CLEAN (TOP & BOTTOM) BEFORE LOADING DO NOT LOAD SLIDES DURING SOFTWARE INITIALIZATION Olympus VS120-L100 Slide Scanner Standard Operating Procedure Startup 1) Red power bar switch (behind monitor) 2) Computer 3) Login: UserVS120 account (no password) 4) Double click: WAIT FOR INITIALIZATION

More information

Brief manual how to start and close the Leica sp2 Confocal. (TCS SP2 AOBS system mounted on a DM IRE2)

Brief manual how to start and close the Leica sp2 Confocal. (TCS SP2 AOBS system mounted on a DM IRE2) Brief manual how to start and close the Leica sp2 Confocal (TCS SP2 AOBS system mounted on a DM IRE2) A. Switching on hardware B. Acquiring and saving images C. Switching off the microscope D. Good working

More information

Nikon Eclipse Ti2-E Widefield/Spinning Disk Confocal Microscope Standard Operation Protocol

Nikon Eclipse Ti2-E Widefield/Spinning Disk Confocal Microscope Standard Operation Protocol Nikon Eclipse Ti-E Widefield/Spinning Disk Confocal Microscope Standard Operation Protocol Please sign on the log sheet before switching on system. Turn on system Turn on A only if confocal mode or laser

More information

Quick Guide for Zeiss 710 Laser Scanning Confocal MGH Cancer Center

Quick Guide for Zeiss 710 Laser Scanning Confocal MGH Cancer Center Quick Guide for Zeiss 710 Laser Scanning Confocal MGH Cancer Center For any questions or concerns, please contact: Linda Nieman lnieman@mgh.harvard.edu Office: (617) 643-9684 Cell: (512) 565-8076 Chenyue

More information

Training Guide for Carl Zeiss AxioZoom V16 Stereo Microscope

Training Guide for Carl Zeiss AxioZoom V16 Stereo Microscope Training Guide for Carl Zeiss AxioZoom V16 Stereo Microscope ZEN 2012 Optical Imaging & Vital Microscopy Core Baylor College of Medicine (2017) Power ON Routine 1 2 If you require fluorescence imaging,

More information

MIF ZEISS VIOLET CONFOCAL ZEN 2009 PROTOCOL

MIF ZEISS VIOLET CONFOCAL ZEN 2009 PROTOCOL MIF ZEISS VIOLET CONFOCAL ZEN 2009 PROTOCOL START-UP On the Switchbox, turn both black switches to the ON position. Wait for the microscope to boot up completely (watch the screen on the side of the microscope).

More information

MetaMorph Imaging Handbook Update 6/4/13

MetaMorph Imaging Handbook Update 6/4/13 MetaMorph Imaging Handbook Update 6/4/13 Startup FIRST turn on mercury lamp (Fluorescence) Computer and monitor Qimaging Camera (on top) Uniblitz Shutters-2 Halogen Lamp (Transmitted Light) Computer Login

More information

Zeiss AxioObserver with ApoTome

Zeiss AxioObserver with ApoTome Zeiss AxioObserver with ApoTome Quick Start User Guide LSU Health Sciences Center-Shreveport Research Core Facility (RCF) Microscopy Table of Contents 1 Start up the system.. Page 3 2 Touch screen controller

More information

Everest System / Slidebook Operating Procedures

Everest System / Slidebook Operating Procedures Everest System / Slidebook Operating Procedures NOTICE: This guide is meant to supplement training, not replace it. All users must be trained first hand by a core employee. Training of others in your lab

More information

Quick Guide for Zeiss 710 Laser Scanning Confocal MGH Cancer Center

Quick Guide for Zeiss 710 Laser Scanning Confocal MGH Cancer Center Quick Guide for Zeiss 710 Laser Scanning Confocal MGH Cancer Center For any questions or concerns, please contact: Linda Nieman lnieman@mgh.harvard.edu Office: (617) 643-9684 Cell: (512) 565-8076 Chenyue

More information

b. Turn the power switch and key to on position for blue laser.

b. Turn the power switch and key to on position for blue laser. OLYMPUS FLUOVIEW 300 CONFOCAL MICOSCOPE OPERATION PROCEDURE 1. Turn ON microscope in this order: 1) Turn on mercury lamp (Note: once the mercury lamp is turned off, DO NOT turn it back on for at least

More information

Imaging Introduction. September 24, 2010

Imaging Introduction. September 24, 2010 Imaging Introduction September 24, 2010 What is a microscope? Merriam-Webster: an optical instrument consisting of a lens or combination of lenses for making enlarged images of minute objects; especially:

More information

DIC Imaging using Laser Scanning Microscopes (LSMs) on Axio Imager Stands

DIC Imaging using Laser Scanning Microscopes (LSMs) on Axio Imager Stands DIC Imaging using Laser Scanning Microscopes (LSMs) on Axio Imager Stands Differential Interference Contrast (DIC) imaging is a technique used to increase contrast in brightfield images. In confocal systems,

More information

LEICA TCS SP5 AOBS TANDEM USER MANUAL

LEICA TCS SP5 AOBS TANDEM USER MANUAL LEICA TCS SP5 AOBS TANDEM USER MANUAL STARTING THE SYSTEM...2 THE LAS AF SOFTWARE...3 THE «ACQUIRE» MENU...5 CHOOSE AND CREATE A SETTING...6 THE CONTROL PANEL...8 THE DMI6000B MICROSCOPE...10 ACQUIRE ONE

More information

Axioscan - Startup. 1. Turn on the Axioscan (button to the left) and turn on the computer. 2. Log on and start the ZEN Blue software from the desktop

Axioscan - Startup. 1. Turn on the Axioscan (button to the left) and turn on the computer. 2. Log on and start the ZEN Blue software from the desktop Axioscan - Startup 1. Turn on the Axioscan (button to the left) and turn on the computer 2. Log on and start the ZEN Blue software from the desktop 3. Press ZEN slidescan and Start System 4. Start by changing

More information

MIF ZEISS LSM510 CONFOCAL USER PROTOCOL

MIF ZEISS LSM510 CONFOCAL USER PROTOCOL MIF ZEISS LSM510 CONFOCAL USER PROTOCOL START-UP Turn on the Mercury Bulb Power Supply (if needed). Power-on the Control Box. Turn on the computer. Open the LSM 510 software. Choose Scan New Images and

More information

Leica Sp5 II Confocal User Guide

Leica Sp5 II Confocal User Guide Leica Sp5 II Confocal User Guide Turning on the Confocal System (instructions are posted in the room) 1. Turn on Laser Power Button 2. Turn Key to On position 3. Turn on Scanner Power Button 4. Turn on

More information

Nasmyth Ultraview Vox User Protocol

Nasmyth Ultraview Vox User Protocol Nasmyth Ultraview Vox User Protocol Switch on all wall sockets labelled Nasmyth, switch camera on (power supply located on table behind monitor), switch on laser switch in laser rack, switch computer on

More information

Zeiss Axioplan 2 imaging microscope and Axiovision software

Zeiss Axioplan 2 imaging microscope and Axiovision software Zeiss Axioplan 2 imaging microscope and Axiovision software Microscopes 1 and 2 in room B501b User Guide Molecular Imaging Unit University of Helsinki www.miu.helsinki.fi 20.5.2010 1 GENERAL... 1 1.1...

More information

LSM 510 Meta Training Notes

LSM 510 Meta Training Notes LSM 510 Meta Training Notes Turning on the system Turn on X-Cite power supply. This supplies light for epifluorescence for viewing your samples through the microscope. Turn on the remote control switch.

More information

Leica DB LB Research microscope and Studo Lite Imaging software

Leica DB LB Research microscope and Studo Lite Imaging software Leica DB LB Research microscope and Studo Lite Imaging software Room B523 User Guide Molecular Imaging Unit University of Helsinki www.miu.helsinki.fi 9.4.2008 1 GENERAL USER INFORMATION... 1 2 SETTINGS

More information

3. are adherent cells (ie. cells in suspension are too far away from the coverslip)

3. are adherent cells (ie. cells in suspension are too far away from the coverslip) Before you begin, make sure your sample... 1. is seeded on #1.5 coverglass (thickness = 0.17) 2. is an aqueous solution (ie. fixed samples mounted on a slide will not work - not enough difference in refractive

More information

Zeiss 880 Training Notes Zen 2.3

Zeiss 880 Training Notes Zen 2.3 Zeiss 880 Training Notes Zen 2.3 1 Turn on the HXP 120V Lamp 2 Turn on Main Power Switch Turn on the Systems PC Switch Turn on the Components Switch. 3 4 5 Turn on the PC and log into your account. Start

More information

Simplified Instructions: Zeiss Brightfield Microscope S1000

Simplified Instructions: Zeiss Brightfield Microscope S1000 Contents General Microscope Set-Up Adjust Illumination Focus Condenser Open Software Image Capture Settings Shading & Color Corrections Image Capture & Viewing Scaling and Measurements Synopsis of Other

More information

Training Guide for Carl Zeiss LSM 5 LIVE Confocal Microscope

Training Guide for Carl Zeiss LSM 5 LIVE Confocal Microscope Training Guide for Carl Zeiss LSM 5 LIVE Confocal Microscope AIM 4.2 Optical Imaging & Vital Microscopy Core Baylor College of Medicine (2017) Power ON Routine 1 2 Verify that main power switches on the

More information

DIC Imaging using Laser Scanning Microscopes (LSM) on Inverted Stands

DIC Imaging using Laser Scanning Microscopes (LSM) on Inverted Stands DIC Imaging using Laser Scanning Microscopes (LSM) on Inverted Stands Differential Interference Contrast (DIC) imaging is a technique used to increase contrast in brightfield images. In confocal systems,

More information

Title: Leica SP5 Confocal User Manual

Title: Leica SP5 Confocal User Manual Title: Leica SP5 Confocal User Manual Date of first issue: 23/10/2015 Date of review: Version: Admin For assistance or to report an issue Office: CG07 or 05 Email: Igmm-imaginghelpdesk@igmm.ed.ac.uk Website:

More information

LSM 780 Confocal Microscope Standard Operation Protocol

LSM 780 Confocal Microscope Standard Operation Protocol LSM 780 Confocal Microscope Standard Operation Protocol Basic Operation Turning on the system 1. Sign on log sheet according to Actual start time 2. Check Compressed Air supply for the air table 3. Switch

More information

Training Guide for Carl Zeiss LSM 510 META Confocal Microscope

Training Guide for Carl Zeiss LSM 510 META Confocal Microscope Training Guide for Carl Zeiss LSM 510 META Confocal Microscope AIM 4.2 Optical Imaging & Vital Microscopy Core Baylor College of Medicine (2017) Power ON Routine 1 2 Turn ON Components and System/PC switches

More information

Training Guide for Carl Zeiss LSM 7 MP Multiphoton Microscope

Training Guide for Carl Zeiss LSM 7 MP Multiphoton Microscope Training Guide for Carl Zeiss LSM 7 MP Multiphoton Microscope ZEN 2009 Optical Imaging & Vital Microscopy Core Baylor College of Medicine (2017) Power ON Routine 1 2 Turn Chameleon TiS laser key from Standby

More information

Dante (Microscope) & Beatrice (Guide) Orth Lab

Dante (Microscope) & Beatrice (Guide) Orth Lab Dante (Microscope) & Beatrice (Guide) Orth Lab Olympus IX81 Widefield Microscope User Guide v. 1.2 (11/2014) Objectives 4x/0.13NA UPLFLN Semi Apo 10x/0.4NA PH UPLAPO Plan Apo 20x/0.8NA PH UPLAPO Plan Apo

More information

CMI STANDARD OPERATING PROCEDURE. Fluoview 300 laser scanning confocal microscope

CMI STANDARD OPERATING PROCEDURE. Fluoview 300 laser scanning confocal microscope CMI STANDARD OPERATING PROCEDURE Fluoview 300 laser scanning confocal microscope CMI documentid:sop001 CONTACT INFORMATION: Peter Owens: 091 494036 (office) Peter.owens@nuigalway.ie Kerry Thompson: 091

More information

DSU Spinning Disk Confocal and Slidebook 4.2 Quick Guide Light Microscopy Core Facility University of Chicago Vers 0.

DSU Spinning Disk Confocal and Slidebook 4.2 Quick Guide Light Microscopy Core Facility University of Chicago Vers 0. DSU Spinning Disk Confocal and Slidebook 4.2 Quick Guide Light Microscopy Core Facility University of Chicago Vers 0.8, February 2007 By: Vytas Bindokas, Ph.D. Core Director 2 The DSU system is optimized

More information

LSM 800 Confocal Microscope Standard Operation Protocol

LSM 800 Confocal Microscope Standard Operation Protocol LSM 800 Confocal Microscope Standard Operation Protocol Turning on the system 1. Switch on the Main switch (labeled 1 and 2 ) mounted on the wall. 2. Turn the Laser Key (labeled 3 ) 90 clockwise for power

More information

Standard Operating Procedure (SOP) for Shared Equipment: Spinning Disk Confocal Microscope

Standard Operating Procedure (SOP) for Shared Equipment: Spinning Disk Confocal Microscope Standard Operating Procedure (SOP) for Shared Equipment: Spinning Disk Confocal Microscope This document is to be used as a supplementary guide and not as a replacement for formal training. DO NOT operate

More information

Training Guide for Leica SP8 Confocal/Multiphoton Microscope

Training Guide for Leica SP8 Confocal/Multiphoton Microscope Training Guide for Leica SP8 Confocal/Multiphoton Microscope LAS AF v3.3 Optical Imaging & Vital Microscopy Core Baylor College of Medicine (2017) Power ON Routine 1 2 Turn ON power switch for epifluorescence

More information

User Manual. Digital Compound Binocular LED Microscope. MicroscopeNet.com

User Manual. Digital Compound Binocular LED Microscope. MicroscopeNet.com User Manual Digital Compound Binocular LED Microscope Model MD82ES10 MicroscopeNet.com Table of Contents i. Caution... 1 ii. Care and Maintenance... 2 1. Components Illustration... 3 2. Installation...

More information

Title: Nikon A1R Confocal User Manual

Title: Nikon A1R Confocal User Manual Title: Nikon A1R Confocal User Manual Date of first issue: 23/10/2015 Date of review: Version: Admin For assistance or to report an issue Office: CG.07 or CG.05 Email: Igmm-imaginghelpdesk@igmm.ed.ac.uk

More information

Leica DMI 4000 tutorial

Leica DMI 4000 tutorial Leica DMI 4000 tutorial Before using the Leica DMI 4000, You will need to put down your name on the reservation system = 1 Welcome to the Leica DMI4000 Microscope tutorial How to start up the system (p.3)

More information

TABLE OF CONTENTS. Safety notes i. Care and Maintenance. ii. 1. Components Illustration Installation of Components.. 4

TABLE OF CONTENTS. Safety notes i. Care and Maintenance. ii. 1. Components Illustration Installation of Components.. 4 TABLE OF CONTENTS Safety notes i Care and Maintenance. ii 1. Components Illustration... 1 2. Installation of Components.. 4 2.1 Installation Diagram... 4 2.2 Installation Procedures 5 3. Operation...11

More information

Microscope ECLIPSE 80i Instructions <Reference>

Microscope ECLIPSE 80i Instructions <Reference> M318E 03.12.CF.1(2/2) Microscope ECLIPSE 80i Instructions Introduction Thank you for purchasing this Nikon product. This instruction manual, which describes basic microscope operations, is

More information

Olympus Time-lapse Microscope Basic operation

Olympus Time-lapse Microscope Basic operation Olympus Time-lapse Microscope Basic operation To start up the microscope 1. Switch on the Olympus UCB. (label as ) 1 Power Switch 2 2. Switch on the MT10. (label as ) Power Switch Page 1 of 18 3 3. Switch

More information

OPERATING INSTRUCTIONS

OPERATING INSTRUCTIONS Zeiss LSM 510 M eta Confocal M icroscope OPERATING INSTRUCTIONS Starting the System: 1. Turn the black knob on the laser box one-quarter turn from Off to On. You will hear the laser cooling mechanisms

More information

Fixed cell DSU Spinning Disk Confocal and Slidebook 5.0 Quick Guide. Light Microscopy Core Facility University of Chicago Vers 1.

Fixed cell DSU Spinning Disk Confocal and Slidebook 5.0 Quick Guide. Light Microscopy Core Facility University of Chicago Vers 1. Fixed cell DSU Spinning Disk Confocal and Slidebook 5.0 Quick Guide Light Microscopy Core Facility University of Chicago Vers 1.0, February 2009 By: Vytas Bindokas, Ph.D., Core Director The fixed cell

More information

Ch. 1 - Installation Guidelines

Ch. 1 - Installation Guidelines Ch. 1 - Installation Guidelines Table of Contents Ch. 1 - Installation Guidelines Introduction... 8 The Image-Pro Driver Interface... 8 Installing the SPOT Image-Pro Driver... 8 Image-Pro Driver Supplement

More information