Imagers- Molecular, Cell Standard Operating Procedures

Transcription

1 Bio-Rad ChemiDoc XRS and Image Lab Software Jump to Export Images to other Apps Floid cell imaging station Life technologies Jump to Chemi-luminescence Protocol Imagers- Molecular, Cell Standard Operating Procedures Prepared by: Bob Morrison STLCC-CPLS, Instrumentation Specialist Initial Jul 10, Revised 24 March 2015 add FLoid Page 1

2 Imager- Molecular : Chemidoc Hood, On/Off, UV Shield Filter Choice Lever: None, Filter 1, Filter 2 USB line to PC MUST be connected to the port as labeled on the PC case. 1. Membrane buttons and LEDs for hood. 2. Normally these are NOT USED as the ImageLab software will control all functions for setup, image capture, and analysis Power lines for the Camera and Imager controls have been routed to the Power strip located right in front of the unit. How to install UV Shield when necessary Pull out drawer, insert in slot at top. Flip the Power Strip to ON to Turn on both of these units Page 2

3 Imager- Molecular : XcitaBlue and White Light Conversion Note: Be sure to run calibration at Edit-Setup with these options once to correct focus and flat field lens operations (Completed in initial setup by BobM 7/12/10) Conversion screen for viewing SYBR Green, SYBR Safe, and other fluorescent gel stains, includes viewing goggles, SYBR Safe filter; for use with Molecular Imager ChemiDoc XRS+ and Gel Doc XR+ systems Page 3

4 Imager- Molecular : ChemiDoc XRS Features The Molecular Imager ChemiDoc XRS+ system is based on CCD high-resolution, high-sensitivity detection technology and modular options to accommodate a wide range of samples and support multiple detection methods including fluorescence, colorimetry, densitometry, chemiluminescence, and chemifluorescence. The system is controlled by Image Lab software to optimize imager performance for fast, integrated, and automated image capture and analysis of various samples. The system accommodates a wide array of samples, from large handcast polyacrylamide gels to small ReadyAgarose gels and various blots. The system is an ideal accompaniment to PCR, purification, and electrophoresis systems, enabling image analysis and documentation of restriction digests, amplified nucleic acids, genetic fingerprinting, RFLPs, and protein purification and characterization. Key benefits include: Gel and blot imaging and analysis are quick and accurate Automated, hands-off routines; no training is required Save and recall all the steps in the workflow for repeatable and reproducible results Optimize the system at setup for image data that is always accurate, reproducible, and free of imaging artifacts Range of applications with special accessories to preserve sample integrity for downstream research while ensuring user safety Comprehensive, automated quantitative analysis of protein and DNA samples in seconds Customize and organize data in reports Obtain publication-quality results quickly Users Manual ChemiDoc XRS and Image Lab Software pdf (2.5mb) Hardware Manual ChemiDoc XRS and Image Lab Software pdf (2.4mb) Page 4

5 Imager- Molecular : ImageLab s/w, Start/Main Page Page 5

6 Imager-Molecular : Imagelab; Quick Start (To take pictures of Ethidium Bromide-Nucleic Acid Gels only) Select New to set up protocol Page 6

7 Imager-Molecular : Imagelab; Quick Start 1. Select Nucleic Acid Gels > Ethidium Bromide 2. Place gel inside imager, click Position Gel, and move gel as needed. Page 7

8 Imager-Molecular : Imagelab; Quick Start Click Run Protocol Page 8

9 Imager-Molecular : Imagelab; Quick Start 1. Save file to USB Drive for further analysis on laptops. 2. Or continue to use analysis tools on current computer Page 9

10 Imager- Molecular : Imagelab, Analysis Tool Box Page 10

11 Imager- Molecular : Imagelab; Protocol Setup, Gel Imaging Page 11

12 Imager- Molecular : Imagelab; Position Gel Page 12

13 Imager- Molecular : Imagelab; Detect Lanes and Bands Page 13

14 Imager- Molecular : Imagelab; Viewing Results Page 14

15 Imager- Molecular: Imagelab; 3-D Projection Page 15

16 Imager- Molecular : ImageLab; Analyze Molecular Weight Page 16

17 Imager- Molecular : Imagelab; Select/Edit Standards Page 17

18 Imager- Molecular : Imagelab; Weight Regression Methods Page 18

19 Imager- Molecular: Imagelab; Lane Profile Option Page 19

20 Imager-Molecular: Chemidoc, Chemiluminescence (by Priya-P. 6/22/11) Select Chemi Set up signal accumulation mode 5 sec to 15 sec with total of 6 images Page 20

21 Imager-Molecular: Chemidoc, Chemiluminescence (by Priya-P. 6/22/11) Move Filter Selection slide bar (behind front panel ) to NO FILTER position Be sure to return Filter Slide Bar to the original setting after this use Page 21

22 Imager-Molecular: Chemidoc, Chemiluminescence Run Protocol Page 22

23 Imager-Molecular: Chemidoc, Chemiluminescence Select 15 sec, then click Yellow Button at left Page 23

24 Imager-Molecular: Chemidoc, Chemiluminescence The results will appear in gray scale. Using comassie or stain free background highlights the chemiluminescence Be sure to return Filter Slide Bar to the original setting after this use Page 24

25 Imager-Molecular: Capture Images for Formatting or Other Documents Use edit features to get image you wish to copy, then Go to Edit in top Toolbar Select Snapshot Page 25

26 Imager-Molecular: Copy Image to Clipboard Select Clipboard (this is the default location for all Windows copy operations) Page 26

27 Imager-Molecular: Open Powerpoint or other App and Paste Clipboard Image to any Slide OR; Press Ctrl (Control) and V keys simultaneously to paste picture from the Clipboard into Powerpoint Page 27

28 Imager-Molecular; Or, Paste Clipboard into MS Word using Ctrl V or Paste icon Page 28

29 Imager- Molecular: Application Tables; Setup Required Page 29

30 Imager- Molecular : Application Tables; Setup Required Page 30

31 Imager- Molecular Imagelab; Standard Curve Page 31

32 Chemiluminescence Yes Colorimetry/densitometry Yes Fluorescence* Gel documentation Yes Yes Hardware Specifications Imager- Molecular: Chemidoc XRS, Specifications Maximum sample size 28 x 36 cm Maximum image area 25 x 26 cm Excitation source Trans-UV and epi-white are standard (302 nm included, with 254 nm and 365 nm available as options); optional trans white, self-powered or conversion screen; optional XcitaBlue UV/blue conversion screen Illumination control 5 modes (trans-uv, epi-white, and no illumination for chemiluminescence are standard); trans white and XcitaBlue conversion screens are optional Detector Supercooled CCD CCD resolution (H x V) 1,392 x 1,040 µm Pixel size (H x V) 6.45 x 6.45 µm Cooling system Peltier Camera cooling temperature 30 C controlled Filter selector 3 positions (2 for filters, 1 without filter, for chemiluminescence) Emission filters 1 included (standard), 3 optional Dynamic range >4.0 orders of magnitude Pixel density (gray levels) 65,535 Dynamic flat fielding Application specific, for all applications Instrument size (L x W x H) 36 x 60 x 96 cm Instrument weight 32 kg Operating Ranges Operating voltage 110/115/230 V AC nominal Operating temperature C (21 C recommended) Operating humidity <70% noncondensing Automation Capabilities Workflow automated selection Application driven, user selected or recalled by a protocol Workflow automated execution Controlled by a protocol via application specific setup for image area, illumination source, filter, analysis, and reporting Workflow reproducibility 100% repeatability via recallable protocols; from image capture to quantitative analysis and reports Autofocus (patent pending) Precalibrated focus for any zoom setting or sample height Image flat fielding** Dynamic; precalibrated and optimized per application Autoexposure 2 user-defined modes (intense or faint bands) *Use the optional XcitaBlue kit if performing SYBR Safe DNA applications, because the UV to blue conversion screen allows you to visualize STLCC_CPLS;Morrison DNA samples 12/18/2015 while protecting against UV damage. Page 32

33 Imager- Molecular : ImageLab S/W Permission to Copy Page 33

34 Imager: Cells, Floid Imaging Station w Printer On/Off toggle, right rear Canon Selphy Printer Hard drive, removable Product plate Info: Page 34

35 Imaging: Cell, Floid Overview The FLoid Cell Imaging Station is an affordable, user-friendly imaging solution for the quick detection and verification of fluorescently-labeled samples. The FLoid Cell Imaging Station captures transmitted light and three-color fluorescent images of your cells and samples right at your benchtop. You do not need to be specially trained or have dedicated staff to operate it. An uncomplicated user interface, streamlined image acquisition process, and real-time, multicolor display allow even imaging novices to produce high-quality images with a few mouse clicks. System specifications include: Objective: Fixed 20X plan fluorite; numerical aperture (NA) = 0.45; working distance (WD) = 5.9 mm. Color Channels: 4 channels (relief phase, blue, green, and red fluorescence) Illumination: LED (50,000 hour life), adjustable intensity Excitation: Blue channel: 390/40 nm; Green channel: 482/18 nm; Red channel: 586/15 nm Emission: Blue channel: 446/33 nm; Green channel: 532/59 nm; Red channel: 646/68 nm Captured Images: 16-bit monochrome, TIFF, PNG, JPG, or BMP ( pixels) LCD display: 15-inch color; display resolution 1366 x 768 pixels; image resolution 1296 x 964 pixels; adjustable tilt Dimensions: (W x H x D): 15.9" x 21.1 x 13.9" / 40.4 cm x 53.6 cm x 35.3 cm; 28 lbs / 12.7 kg Hot link to Floid Users Manual (Life Technologies pdf (2.5mb) Page 35

36 Imager; Cell, Floid, Quick Start Guide Page 1 Page 36

37 Imager; Cell, Floid, Quick Start Guide pg 2 Page 37

38 : Other Options Future home of other detailed procedures or printout options. Page 38