Simplified Instructions: Olympus Widefield Microscope S1230

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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 Simplified Instructions: Olympus Widefield Microscope S1230 This is an Overview Only intended to refresh specific items learned in the Hands-On Training Use only in conjunction with your own much more detailed notes taken during your training

1, Reserve Time on Scheduler 2, Remove Dust Cover Please do NOT place on the table beside the camera. Use shelf or drawers.

3, Power-Up Locate Lamp Power Supply (to Left of System) Ensure Power to All Other Components is Off Before Turning Lamp On

3, Power-Up Turn on Power to Lamp (Depress Top of Red Button)

3, Power-Up Ignite Lamp (Lift Toggle to Manual, Release) please do not set toggle to Auto Auto decreases lamp life

3, Power-Up Locate Power Conditioner for Other Components on Upper Shelf

3, Power-Up Turn on Power to Other Components Please Do NOT Touch the Individual Power Switches For these Components

3, Power-Up Turn on Computer Computer is Under Monitor

4, Access Software Use Windows Log-In/Password provided to you at training Click on Meta Imaging Series icon (some may have a direct short cut to Metamorph on their Desktop)

5, Open Metamorph (double-click icon) (may be on desktop for some)

6, Viewing Sample Move Desired Objective into Position

6, Viewing Sample Indicate Objective in Metamorph Using pull-down menu (important for correct scale-bar after images collected)

6, Viewing Sample Clean Objective

6, Viewing Sample Direct Emissions to Ocular Viewing

6, Viewing Sample For Brightfield Viewing, Turn on halogen lamp

6, Viewing Sample For Brightfield Viewing, Turn on halogen lamp Filters on top Condensor

6, Viewing Sample For Brightfield Viewing, Slider changes intensity of halogen lamp

6, Viewing Sample For Fluorescence Viewing Turn off Halogen Lamp Indicate Filter Set using pull-down menu Note Mirror information ( m3 etc.)

6, Viewing Sample For Fluorescence Viewing Move Correct Mirror into Position (mechanical shutter must be open)

6, Viewing Sample For Fluorescence Viewing Toggle Shutter On/Off

6, Viewing Sample For Fluorescence Viewing Focus using indicated knob ONLY (Do NOT use knobs on microscope) Objective Up (toward slide) is clockwise

6, Viewing Sample For Fluorescence Viewing Toggle between Course (default), medium and fine focus

Remember to keep exposure to a minimum to prevent photobleaching i.e. keep shutter off as much as possible 6, Viewing Sample For Fluorescence Viewing Move Stage with joystick left toggle switch is for course to fine (default) stage movements

7, Simple Image Capture Direct Emissions to Camera ( Side Port, SP)

7, Simple Image Capture Under Acquire Pull-Down, Open Acquire Control (this control sets standard image collection parameters)

7, Simple Image Capture In Acquire Control, Display Set Exposure Time NEVER change anything except Items discussed below!!!

7, Simple Image Capture In Acquire Control, Display Set Binning (note, can set Live binning differently)

7, Simple Image Capture In Acquire Control, Display Set intensity where white is for viewing (preference is 4095) (can use autoscale also) NEVER change image gamma from 1

7, Simple Image Capture In Acquire Control, Special Set Gain Do NOT change anything else

7, Simple Image Capture In Acquire Control Click Show Live to Start Image Capture Keep on Full Chip, there is no reason to collect smaller size

7, Simple Image Capture Use continuously update Live image to focus your image on the screen (change exposure times accordingly) Stop Live toggle shuts down screen

7, Simple Image Capture Icons on Show Live (or Captured Image) Set Display Size, Color or Display Intensity

7, Simple Image Capture Click Acquire to Capture Image Exactly as Seen Under Show Live

8, Image Saving Under File : Save Save Your Captured Image to Your File on the Server

8, Image Saving To Save a Series with Same Base Name a, Open Setup Sequential File Names

8, Image Saving To Save a Series with Same Base Name b, Type In Base Name

8, Image Saving To Save a Series with Same Base Name c, Set Number of Digits in Name (image number is starting number) (output style example is shown)

8, Image Saving To Save a Series with Same Base Name d, Set Image Type Strongly recommend Metamorph TIFF for single images (grouped images- stacks-discussed later)

8, Image Saving To Save a Series with Same Base Name e, Select Server and Folder Location (can create a new Folder in location with New ) Click OK to finish Naming Set-up

8, Image Saving To Save a Series with Same Base Name f, Save Using Sequential File Name Saves files to designated location with Name 01, 02, 03, etc.

9, Multi-Image Capture Set Basic Image Capture Parameter Using the Acquire Box Open Acquire Multiple Wavelengths

9, Multi-Image Capture Set the Number of Different Wavelengths to Capture

9, Multi-Image Capture For Each Wavelength, Indicate Filter Set (Note: You MUST Choose Compatible Wavelengths that Use a Single Mirror)

9, Multi-Image Capture Manually Position the Correct Mirror Enter Exposure Times (Usually Determined Using Show Live) (Can Also Click Acquire to See Test Image for Each Wavelength)

9, Multi-Image Capture Click Top Acquire to Capture All Images

9, Multi-Image Capture Toggle Between Images in the Stack Using the Arrows. Other Image Display Consideration As Described under 7, Simple Image Capture

9, Multi-Image Capture Saving As Described under 7, Simple Image Capture Except That Save as a Metamorph Stack

10, Capture a Z-Series Open Dialog Under Acquire Again, Basic Parameters Set Under Acquire Drop-Down

10, Capture a Z-Series Set a, Number of Z-sections to collect

10, Capture a Z-Series Set b, Number of Steps between Each Z-section (positive steps move from bottom to top)

10, Capture a Z-Series Set c, to start where last focused i.e. Current Position Others are set positions on stage: Do NOT use those

10, Capture a Z-Series Set d, Planes x Spacing Do NOT Use Others

10, Capture a Z-Series Set e, to Collect as a Stack using f, the Desired Filter Set (Use Correct Mirror) F, Exposure Time Set Under Acquire

10, Capture a Z-Series g, OK Captures Z-series

10, Capture a Z-Series Toggle Between Images in the Stack Using the Arrows. Other Image Display Consideration As Described under 7, Simple Image Capture

10, Capture a Z-Series Saving As Described under 7, Simple Image Capture Except That Save as a Metamorph Stack

11, Capture a Timelapse Open Dialog Under Acquire Again, Basic Parameters Set Under Acquire Drop-Down

11, Capture a Timelapse a, Set Time Between Start of Each Collection (If Duration of Collection Longer than Time, It Is Not Permissible)

11, Capture a Timelapse b, Set Number of Timepoints to Collect c, Set Filters with Correct Mirror d, Select Exposure Time e, Click OK to Start Collection f, Save as a Stack

12, Combining Any of Multi-λ, Z or Time-Series Open Apps: Multi Dimensional Acq.

12, Combining Any of Multi-λ, Z or Time-Series a, Under Main, Check Types of Collections to Combine (Appropriate Interface Boxes Appear)

12, Combining Any of Multi-λ, Z or Time-Series This Dialog Contains Parameters Also Found Under Acquire (But Not Gain) (Recommend to Set Under Acquire And Let Auto-populate)

12, Combining Any of Multi-λ, Z or Time-Series b, Collections Under This Dialog Are Saved With the Name Specified in the Specified Folder

12, Combining Any of Multi-λ, Z or Time-Series c, Enter Number of Wavelengths and Exposure Time for Each Wavelength

12, Combining Any of Multi-λ, Z or Time-Series d, Enter Z-Series Settings (Multiple Stage Positions in Main Menu can be use to set multiple x,y positions)

12, Combining Any of Multi-λ, Z or Time-Series e, Enter Timelapse Settings (Minimum Time Interval Determined by λ, x-y and Z-Settings)

12, Combining Any of Multi-λ, Z or Time-Series f, Acquire All Settings

12, Combining Any of Multi-λ, Z or Time-Series g, Use Review Multi Dim. Data To See Results

There are a Wide Variety of Functions Available in Metamorph Not Detailed In This Short Synopsis Applications Under File Shown Here

There are a Wide Variety of Functions Available in Metamorph Not Detailed In This Short Synopsis Applications Under Edit Shown Here

There are a Wide Variety of Functions Available in Metamorph Not Detailed In This Short Synopsis Applications Under Regions Useful, But Not Shown Here

There are a Wide Variety of Functions Available in Metamorph Not Detailed In This Short Synopsis Applications Under Stack Shown Here

There are a Wide Variety of Functions Available in Metamorph Not Detailed In This Short Synopsis Applications Under Device Shown Here UNDER NOT CIRCUMSTANCE DOES ANYBODY TOUCH CONFIGURE ILLUMINATION OR MAGNIFICATION. DON T EVEN LOOK AT THOSE

There are a Wide Variety of Functions Available in Metamorph Not Detailed In This Short Synopsis Applications Under Device Shown Here Can Use Stage to Mark Stage Positions

There are a Wide Variety of Functions Available in Metamorph Not Detailed In This Short Synopsis Applications Under Display Shown Here Scale Image is Very Useful to Change Display Settings to a Consistent Level

There are a Wide Variety of Functions Available in Metamorph Not Detailed In This Short Synopsis Applications Under Display Shown Here Overlay Image Combine Stacks Into an Overlaid Image

There are a Wide Variety of Functions Available in Metamorph Not Detailed In This Short Synopsis Applications Under Process Shown Here Image Manipulation, Such As Background Subtraction and Much More

There are a Wide Variety of Functions Available in Metamorph Not Detailed In This Short Synopsis Applications Under Log Shown Here Log is Used to Record Data Into An Excel Sheet

There are a Wide Variety of Functions Available in Metamorph Not Detailed In This Short Synopsis Applications Under Measure Shown Here

There are a Wide Variety of Functions Available in Metamorph Not Detailed In This Short Synopsis Applications Under Apps Shown Here

There are a Wide Variety of Functions Available in Metamorph Not Detailed In This Short Synopsis Applications Under Journal Shown Here Journals are Very Useful For Combining Applications Seemlessly Together

There are a Wide Variety of Functions Available in Metamorph Not Detailed In This Short Synopsis Use Window to Find Open Windows

There are a Wide Variety of Functions Available in Metamorph Not Detailed In This Short Synopsis Go to Help to Obtain Instructions for Any of The Prior Functions