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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. 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)
Ex396/Em580	387_11nm	607_36nm	Quadband (TRITC-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
Ex488/Em617	485_20nm	607_36nm	Quadband (TRITC-FIXED)
Ex510/Em580	485_20nm	607_36nm	Quadband (TRITC-FIXED)
AlexaF594 (put HcRED_LIVE cube in Pos. 5)	560_25nm	632_22nm	Triband
FRET YFP-ALX594 (put HcRED_LIVE cube in Pos. 5)	504_12nm	632_22nm	Triband
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 ASCII Data ASCII Data ASCII Data Exciter 2 ASCII Data Exciter 3 ASCII Data
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 ASCII Data ASCII Data ASCII Data Exciter 2 ASCII Data Exciter 3 ASCII Data Exciter 4 ASCII Data
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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