BD LSR II

BD LSR II - A
BD LSR II - B
Controls required for multiparametric analysis BD LSR II - A

BD LSR II - A

Blue laser 488nm
Red laser 640nm
Violet laser 405nm
Yellow-green laser 561nm

The BD LSRII-A is configured to simultaneously detect up to 14 fluorochromes as well as cell or particle size and granularity (FSC and SSC).

All acquisitions are digitized to enhance resolution, detection and measurement of fluorescence.

Usable colours

Lasers	Detectors	Filters	Fluochromes
Blue 488nm	FSC		Forward Scatter
Blue 488nm	SSC	488/10	Side Scatter
Blue 488nm	B	530/30	FITC, CFSE GFP, Alexa-488
Blue 488nm	A	710/20	PerCP-Cy.5
Red 640 nm	C	660/20	APC, Alexa-647
Red 640 nm	B	720/30	APC-700, Alexa-680
Red 640 nm	A	780/60	APC-Cy7, APC-H7
Violet 405nm	D	440/40	Pacific Blue, V450 DAPI, Alexa-405
Violet 405nm	C	525/50	AmCyan, V500, Alexa-430, Aqua Vivid
Violet 405nm	B	610/20	Qdot -605
Violet 405nm	A	780/40	Qdot - 800
Yellow-Green 561 nm	E	582/15	PE
Yellow-Green 561 nm	D	610/20	PE-TxRed, ECD, PI
Yellow-Green 561 nm	C	670/40	PE-Cy5
Yellow-Green 561 nm	B	710/50	PE-Cy5.5
Yellow-Green 561 nm	A	780/60	PE-Cy7

For the detection of other fluorochromes than those mentioned above, to request a training session, or for other information, please contact the facility manager by email: cytometrie.crchum@ssss.gouv.qc.ca

BD LSR II - B

Blue laser 488nm
Red laser 633nm
Violet laser 407nm

The BD LSRII-A is configured to simultaneously detect up to 10 fluorochromes as well as cell or particle size and granularity (FSC and SSC).

All acquisitions are digitized to enhance resolution, detection and measurement of fluorescence.

Usable colours

Lasers	Detectors	Filters	Fluorochromes
Blue 488nm	FSC		Forward Scatter
Blue 488nm	SSC	488/10	Side Scatter
Blue 488nm	E	530/30	FITC, CFSE GFP, Alexa-488
Blue 488nm	D	575/26	PE-PI
Blue 488nm	C	610/20	PE-TxR
Blue 488nm	B	670/14	PE-Cy5, PerCP
Blue 488nm	A	780/40	PE-Cy7
Red 633nm	C	660/20	APC, Cy5, Alexa-647
Red 633nm	B	720/30	APC-Cy5.5, Alexa-680, 700
Red 633nm	A	780/60	APC-Cy7
Violet 407nm	B	440/40	Alexa-405, Pacific Blue, Cascade Blue
Violet 407nm	A	525/50	Alexa-430, CFP, AMCyan

Required controls for multiparametric analysis

Compensation controls

The BD LSRII has automatic compensation settings.

To make optimal use of this function, it is essential to prepare a series of controls, such as:

unstained cells, with the same autofluorescence as your experimental sample;
cells labelled with each fluorochrome used in your experiment;
if you do not wish to use your samples, BD CompBeads can be used with the same antibodies as those in your experimental sample; using them will increase brightness.

Fluorescence minus one controls (FMOC) for gating

In multicolor cytometry, accurate gating requires the use of fluorescence Minus one controls

FMOC allow you to place your gates strategically and rapidly and permit analysis of wekly positive cells populations.

For further information, please contact the facility manager by email: cytometrie.crchum@ssss.gouv.qc.ca

Isotypic controls

These controls permit evaluation of the specificity of your conjugated antibodies.

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