BCH FICR-PCMM Flow and Imaging Cytometry Resource

 

New FICR Facility Policies in place due to Covid-19 Restrictions

Facility access will be dictated by university/hospital mandated building closures for daily disinfection/cleaning.

Mandatory PPE, social distancing and new disinfection policies will be in effect throughout the facility.

Instrument Training will be suspended until social distancing guidelines are relaxed.

NO WALK-UP INSTRUMENT USE WILL BE PERMITTED; all use must be reserved in iLab prior to the appointment time so it can be approved by core staff.

Existing users will reserve instruments through iLab as usual, but all reservations will require core approval, so advance planning is required.

 

If you are in need of assisted sorting or analysis, please contact Viraga.haridas@childrens.harvard.edu 

Welcome

Welcome to the Flow and Imaging Cytometry Resource (FICR) at Boston Children's Hospital. 

FICR provides a platform and instrumentation for researchers interested in using flow and imaging cytometry, as well as molecular imaging for their biological and biomedical research questions.

FICR provides scientists with access to a number of critical technologies including Flow Cytometry and Cell Sorting, Imaging Cytometry and In Vivo Molecular Imaging.  

 

Leadership

Viraga Haridas, Ph.D; Core Director, Flow and Imaging Cytometry Resource

James V. Falvo, Ph.D.; Scientific Administrative Director, Program in Cellular and Molecular Medicine 

 

Location and Hours of Operation

Hours	Locations
Open All Hours	Armenise Building (200 Longwood Ave): AR-231 and AR-239 Center for Life Sciences (3 Blackfan Cr): CL-03034 and CL-3137 New Research Building: Basement NRB 0044

Billing Policies

Users will be billed for the time they reserve on the iLab calendar OR the actual instrument time used, whichever is greater. 

There is a 24-hour cancellation policy. Users that do not cancel or show up for their reservations will be billed for the time reserved, unless they contact FICR staff in advance of their scheduled appointment time to cancel or modify the reservation. All scheduling changes requested within 24 hours of the reserved time will be made at the discretion of FICR staff.

Users that reserve instruments in the facility must abide by the policies set forth by the facility.

 

Resource Information

FACS Cell Sorters and Analyzers (Self operated after training)

 

SONY SH-800Z: 4-Laser, 6-Color . Warren Alpert building (WA-138) and Center for Life Sciences Building (CL-3117)

 

Lasers:  488 nm, 405 nm, 561 nm and 638 nm

 

Filters:                                                                                   

450/50: PacBlue/DAPI (FL1)

525/50: FITC/GFP (FL2)                                                          

585/15: PE (FL3)                                        

665/30: APC (FL4)

720/60: AF700/PCP-Cy5.5 (FL5)                                                                       

785/60: PE-Cy7/APC-Cy7 (FL6)

 

FACSARIA II SORP: 5-Laser, 16-Color. Armenise building (AR-239)

     

488 nm                                406 nm                               592 nm                                 640 nm                         355 nm

530/30: FITC                       450/50: PacBlue                630/20: mCherry                    660/20: APC                 383/17: BUV395

575/25: PE                          530/30: AmCyan                670/20: DsRed (APC)            720/40: AF700              530/30: Indo-1

605/12: PE-Cy5                   605/12: QDot605                                                          780/60: APC-Cy7

670/30: PCP-Cy5.5              705/70: QDot705

780/60: PE-Cy7

     Cytek Aurora: 4-Laser, 48-Florescence Parameters, 3 Scatter Channels. Armenise building (AR-239) 

        

          Lasers:  405 nm, 488 nm, 561 nm and 640 nm

          Runs 5ml tubes, Automated Sample Loader (ASL) which supports a variety of 96-well plates. 

                              405nm                                                                    488nm                                                               561nm                                                                     640nm

        Channel   Bandpass   Wavelength Range          Channel   Bandpass   Wavelength Range          Channel   Bandpass   Wavelength Range               Channel   Bandpass   Wavelength Range 

              V1        428/15       420-435                                B1          508/20       498-518                                YG1         577/20       567-587                                    R1         661/17        653-670 

              V2        443/15       436-451                                B2          525/17       516-533                                YG2         598/20       588-608                                    R2         679/18        670-688 

              V3        458/15       451-466                                B3          542/17       533-550                                YG3         615/20       605-625                                    R3         697/19        688-707 

              V4        473/15       466-481                                B4          581/19       571-590                                YG4         661/17       653-670                                    R4         717/20        707-727 

              V5        508/20       498-518                                B5          598/20       588-608                                YG5         679/18       670-688                                    R5         738/21        728-749 

              V6        525/17       516-533                                B6          615/20       605-625                                YG6         697/19       688-707                                    R6         760/23        749-772 

              V7        542/17       533-550                                B7          661/17       653-670                                YG7         720/29       706-735                                    R7         783/23        7 72-795 

              V8        581/19       571-590                                B8          679/18       670-688                                YG8         750/30       735-765                                    R8         812/34        795-829

              V9        598/20       588-608                                B9          697/19       688-707                                YG9         780/30       765-795 

              V10      615/20       605-625                                B10         717/20      707-727                                YG10       812/34       795-829

              V11      664/27       651-678                                B11         738/21      728-749 

              V12      692/28       678-706                                B12         760/23      749-772 

              V13      720/29       706-735                                B13         783/23      772-795 

              V14      750/30       735-765                                B14         812/34      795-829

              V15      780/30       765-795

              V16      812/34       795-829

 

 

FACSCanto II-AR: 3-Laser, 8-Color. Armenise building (AR-239)

         

488 nm                                405 nm                              640 nm                       

530/30: FITC                       450/50: PacBlue                660/20: APC                  

562/42: PE                          510/50: AmCyan               780/60: APC-Cy7

670 LP: PCP/PCP-Cy5.5                                                                        

780/60: PE-Cy7

 

FACSCanto II-CLSB: 3-Laser, 8-Color. 3-Blackfan Circle (CLSB 3^rd Floor, CL-03034) With Plate loader

      

 

488 nm                                405 nm                              640 nm                       

530/30: FITC                       450/50: PacBlue                660/20: APC                  

562/42: PE                          510/50: AmCyan               780/60: APC-Cy7

670 LP: PCP/PCP-Cy5.5                                                                        

780/60: PE-Cy7

 

Hybrid "In Flow" Microscope - ImageStream^X Mark II (self-operated after training).

Armenise building (AR-231)

 

Imaging flow Cytometry with Hybrid "In Flow" Microscope	Optical Configuration and Parameters	Applications
Quantitative imaging flow cytometry (IFC) allows to analyze cells, cellular clusters,aggregates, and extracellular vesicles with sensitivity higher than conventional flow cytometer	120 mW 405nm laser 200 mW 488 nm laser, 200 mW 561 nm laser, 150 mW, 658 nm laser  Side scatter, 2 Brightfield, up to 9 fluorescent channels, 2 CCD cameras speed:up to 5,000 events/sec; up to 100,000 images per file Microscope objectives: x20, x40, x60	CELL SIGNALING; Nucleo-Cytoplasmic Translocation; INTERNALIZATION, Co-localisation, Cell-Cell Interaction;  APOPTOSIS; DNA Damage and Repair; Cell Cycle and Mitosis; SPOT COUNTING; Morphology; CELL SHAPE; EXOSOMES More than 100 morphological and fluorescent features with >20 masks; more than 2000 parameters - Feature Finder-automatically defines statistically significant features between two different panels of images picked by investigator 1 dot on dotplot is associated with up to 12 brightfield and fluorescent images

 

Molecular Imaging In Vivo/ Pre-clinical in vivo imaging (self-operated after training).

Location: New Research Building, Basement.

  

 

Advanced optical imaging	Optical Configuration	Applications
The fluorescent and bioluminescent reporters	28 efficiency filters spanning 430-850 nm; CCD camera: 13.5 micron pixels, 2048x2048	Non-invasive longitudinal monitoring of disease progression, cell trafficking and gene expression patterns in living animals
Epi-illumination  Trans-illumination	High sensitivity in vivo fluorescence and bioluminescence imaging; High resolution: to 20 microns with 3.9 cm field of view	Multi-spectral imaging with superior spectral unmixing; capable to distinguish multiple bioluminescent and fluorescent reporters

 

 

Contacts

Name	Role	Phone	Email	Location
Viraga Haridas	Core Director	339-970-1931	viraga.haridas@childrens.harvard.edu	ARM 239