Standard Operating Procedures - STLCC.edu

STLCC_CPLS;Morrison 12/18/2015

Imagers- Molecular, Cell Standard Operating Procedures

Prepared by: Bob Morrison

STLCC-CPLS, Instrumentation Specialist

Initial Jul 10, Revised 24 March 2015 add FLoid

Bio-Rad ChemiDoc XRS

and Image Lab Software

Jump to Chemi-luminescence Protocol

Jump to Export Images to other Apps

Floid cell imaging station

Life technologies

Imager- Molecular : Chemidoc Hood, On/Off , UV Shield


Power lines for the Camera

and Imager controls have

been routed to the Power

strip located right in front

of the unit.

Flip the Power Strip to “ON”

to Turn on both of these

units

How to install UV Shield when necessary

Pull out drawer, insert in slot at top .

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

Filter Choice Lever:

None, Filter 1, Filter 2

USB line to PC MUST

be connected to the

port as labeled on the

PC case.

Imager- Molecular : XcitaBlue and White Light Conversion


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

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)

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)

users.stlcc.edu/Departments/fvbio/Imager_Molecular_ChemidocXRS_Imagelabsw.pdf

users.stlcc.edu/Departments/fvbio/Imager_Molecular_ChemidocXRS_Hardware_manual.pdf

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


Imager-Molecular : Imagelab; Quick Start(To take pictures of Ethidium Bromide-Nucleic Acid Gels only)


Select “New…” to

set up protocol

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.



Click “Run Protocol”



1. Save file to USB

Drive for further

analysis on laptops.

2. Or continue to use

analysis tools on

current computer

Imager- Molecular : Imagelab, Analysis Tool Box


Imager- Molecular : Imagelab; Protocol Setup, Gel Imaging



Imager-

Molecular :

Imagelab;

Position Gel

Imager- Molecular :

Imagelab; Detect

Lanes and Bands


Imager- Molecular : Imagelab; Viewing Results


Imager- Molecular: Imagelab; 3-D Projection



Imager-

Molecular :

ImageLab;

Analyze

Molecular

Weight


Imager- Molecular : Imagelab; Select/Edit Standards


Imager- Molecular

: Imagelab; Weight

Regression

Methods

Imager- Molecular: Imagelab; Lane Profile Option


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

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

Imager-Molecular: Chemidoc, Chemiluminescence


Run Protocol



Select 15 sec, then click Yellow Button

at left



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

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

Imager-Molecular: Copy Image to Clipboard


Select Clipboard (this is

the default location for all

Windows copy operations)

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

Imager-Molecular; Or, Paste Clipboard into MS Word using

Ctrl –V or Paste icon



Imager-

Molecular:

Application

Tables; Setup

Required


Imager- Molecular : Application Tables; Setup Required


Imager- Molecular

Imagelab; Standard

Curve

Chemiluminescence Yes

Fluorescence* Yes

Colorimetry/densitometry Yes

Gel documentation Yes

Hardware 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 10–28°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 DNA samples while protecting against UV damage.

Imager- Molecular: Chemidoc

XRS, Specifications


Imager- Molecular : ImageLab S/W Permission to Copy


Imager: Cells, Floid Imaging Station w Printer


On/Off toggle, right rear

Hard drive, removable

Product plate Info:

Canon Selphy Printer

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 (1296 × 964 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)

users.stlcc.edu/Departments/fvbio/Imaging_floid_cell_imaging_station_manual.pdf

Imager; Cell, Floid, Quick Start Guide Page 1


Imager; Cell, Floid, Quick Start Guide pg 2



: Other Options

• Future home of other detailed procedures or printout options ….

Standard Operating Procedures - STLCC.edu

Documents

Transcript of Standard Operating Procedures - STLCC.edu