High-Throughput Bioscience Center (HTBC)

High-Content Imaging

The Molecular Devices ImageXpress Micro (located in CCSR 0116)

Please contact us for key card access to this room. ***Now upgraded with bright field/phase contrast and robotic plate handling.***
iLabs IXMicro Schedule
View only Calendar for non-registered users

For high-content automated cellular imaging and analysis we are utilizing the Molecular Devices ImageXpress Micro (datasheet IXMicroXL) with the live cell option. We use Semrock filters. See below for our filter configurations.

The IXMicro contains a fully automated inverted epifluorescence microscope that is capable of scanning a 6-, 24-, 96- or 384-well microplate and recording multiple fluorescent images in a few minutes. The ImageXpress Micro is then able to quantify morphological properties through the MetaXpress analysis software (modules). Our current ImageXpress Micro configuration permits the visualization of both fixed cells and live cells, as the instrument has a temperature-, humidity-, and carbon dioxide-controlled chamber. We have added the phase contrast/brightfield option and also have integrated an external automated incubator (Thermo Cytomat 2C, 42 plates, Datasheet) to permit long-term or large-scale studies of cellular events, and a robotic plate stacker (Thermo CataLyst Express Microplate Handler, Datasheet, System, Polara Scheduling Software, Datasheet) thereby increasing the throughput of this workstation.

ImageXpress Micro iLabs IXMicro Schedule, Analysis Computer1, Analysis Computer2

To use the ImageXpress Micro, please contact Jason Wu (x5-6004).

MetaXpress 5 Applications Modules:
Angiogenesis tube formation Cell health
Cell scoring Count nuclei
Granularity Live/dead
Mitotic Index Neurite outgrowth
Transfluor® Translocation — basic and enhanced

Preferred Imaging Plates
Wells
Vendor
Catalog Number
Objectives
24
Costar
 
All
96
Greiner
655090
All
96
Costar*
3904
All
384
Greiner
781091
All
384
Costar*
3712
All
 
*Secondary Preference
   

Technical Information
Objectives Excitation Filters Emission Filters Dichroics
4X 0.2 S Fluor 340_26nm CUBE 1-Quad 410/504/582/669 Quadband1
10X 0.3 Plan Fluor 427_10nm 440_40/521_21/607_34/700_45 410/504/582/669
20X 0.45 Plan Fluor ELWD 504_12nm CUBE 2-triple 464/542/639 Triband
40X 0.6 Plan Fluor ELWD 589_15nm 464_23/542_27/639_42 444/521/608
  387_11nm CUBE 3-CFP-LIVE  
Phase Objectives 485_20nm 483_32 458
4X PhL Plan Fluor DL 0.13 NA 560_25nm CUBE 4-YFP-LIVE  
10X Ph1 Plan Fluor DL 0.30 650_13nm 542-27 520
20X Ph1 Plan Fluor ELWD DM   CUBE 5-HcRed-LIVE  
    632-22 Triband
FIXED Cells Profile   CUBE 6-DAPI-FIXED (Pos. 3)  
    440-40 409
    CUBE 7-FITC-FIXED (Pos. 4)  
    525-30 506
    CUBE 8-TRITC-FIXED (Pos. 5)  
    607-36 Quadband
FRET Profile   CUBE 9-YFP-FRET (Pos. 4)  
(CFP-Live in Pos. 3)   542-27 458
    CUBE 10-Fura-2 (Pos. 5)  
    510-84nm Fura-2 (409)
Cy5 (Dedicated)   CUBE 11-Cy5 (Pos. 1)  
    692_40nm 660

Filter Sets
Label Name Excitation Emission Dichroic
Fura-2 340x 340_26nm D510-80nm/Fura-2 Fura-2
Fura-2 387x 387_11nm D510-80nm/Fura-2 Fura-2
CCF4 (FIXED) 387_11nm 525_30nm 506 (FITC-FIXED)
CFP 427_10nm 464_23/542_27/639_42 Triband
CFP-LIVE 427_10nm 483_32nm 458
FRET-LIVE 427_10nm 542_27nm 520 (YFP-LIVE)
YFP-FRET 427_10nm 542_27nm 458
YFP 504_12nm 464_23/542_27/639_42 Triband
YFP-LIVE 504_12nm 542_27nm 520
HcRED 589_15nm 464_23/542_27/639_42 Triband
HcRED-LIVE 589_15nm 632_22nm Triband
DAPI 387_11nm 440_40/521_21/607_34/700_45 Quadband
DAPI-FIXED 387_11nm 440_40nm 409
FITC 485_20nm 440_40/521_21/607_34/700_45 Quadband
FITC-FIXED 485_20nm 525_30nm 506
TRITC 560_25nm 440_40/521_21/607_34/700_45 Quadband
TRITC-FIXED 560_25nm 607_36nm Quadband
Cy5 650_13nm 440_40/521_21/607_34/700_45 Quadband
Cy5 650_13nm 692_40nm 660

 
 
CFP/YFP/HcRed-3X-A Triple-band "Pinkel" Filter Set
 
 Optimized for these fluorophores
Cyan: CFP, AmCyan, SYTOX Blue, BOBO-1, BO-PRO-1
Yellow: YFP, Calcium Green-1, Fluo-3, Rhodamine 123
Red: HcRed, Cy3.5, Texas Red®, Alexa Fluor 594
Complete list of fluorophores for this filter set
 
Actual measured data from typical filters
     
 
Exciter 1
Dichroic
Emitter
Exciter 2
Exciter 3
 
 
 Set description
This 5-filter "Pinkel" set is perfectly optimized for imaging CFP, YFP, and HcRed with high brightness, extremely low crosstalk, and superb signal-to-noise ratio. The set is also ideal for samples triple-labeled with CFP, YFP, and an orange or red dye, like Texas Red. It is comprised of three single-band exciters along with a triple-band beamsplitter and a triple-band emitter, and thus works best with an exciter filter wheel or slider.
 
Filter Average Transmission Center Wavelengths Bandwidths *
 Exciter 1 > 90% 427 nm 10 nm
 Exciter 2 > 90% 504 nm 12 nm
 Exciter 3 > 90% 589 nm 15 nm
 Emitter

> 90%, 90%, 90%

464, 542, 639 nm 23, 27, 42 nm
  Average Reflection Reflection Bands Transmission Bands
 Dichroic > 95%
> 95%
> 95%
420 - 430 nm
496 - 510 nm
579 - 596 nm
451 - 480 nm
530 - 561 nm
618 - 664 nm
 

 
 
DA/FI/TR/Cy5-4X-A Quad-band "Pinkel" Filter Set
 
 Optimized for these fluorophores
Blue: DAPI, Hoechst, AMCA, Alexa Fluor 350
Green: FITC, GFP, rsGFP, BODIPY, Alexa Fluor 488        
Orange: TRITC, Cy3™, Texas Red®, MitoTracker Red,
  Alexa Fluor 568 & 594
Red: Cy5™
, APC, TOTO-3, TO-PRO-3, Alexa Fluor 647 & 660
Complete list of fluorophores for this filter set
 
Actual measured data from typical filters
     
 
Exciter 1
Dichroic
Emitter
Exciter 2
Exciter 3
Exciter 4
 

 
 Set description
This 6-filter "Pinkel" quad-band set is designed for sequentially imaging DAPI, FITC, TRITC, and Cy5 (and other key fluorophores – see complete list above). This state-of-the-art set represents a break-through in quad-band performance, and has been tested by independent researchers to have up to four times the brightness, twice the signal-to-noise ratio and substantially lower crosstalk than any other such quad-band filter set on the market. The complete set is comprised of a quad-band beamsplitter and a quad-band emitter (these can be offered assembled in a microscope cube), plus four “no burn-out” single-band exciters that are intended to be mounted in an exciter filter wheel or slider.
 
Filter Average Transmission Center Wavelengths Bandwidths *
 Exciter 1 > 85% 387 nm 11 nm
 Exciter 2 > 90% 485 nm 20 nm
 Exciter 3 > 90% 560 nm 25 nm
 Exciter 4 > 90% 650 nm 13 nm
 Emitter

> 90%, 90%,
90%, 90%

440, 521,
607, 700 nm
40, 21,
34, 45 nm
  Average Reflection Reflection Bands Transmission Bands
 Dichroic > 95%
> 95%
> 95%
> 95%
381 - 392 nm
475 - 495 nm
547 - 572 nm
643 - 656 nm
420 - 460 nm
510 - 531 nm
589 - 623 nm
677 - 722 nm
 

Old ImageXpress 5000A Webpage

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