CRF Intranet

Flow Cytometer Facility (KSBS Shared IIT Delhi)

A flow cytometer provides multi-parametric analysis of single cells/particles in suspension. The presence of three lasers (488, 562, 632 nm) allows measurement of fluorescent properties of the cells/particles. Using the above lasers, it is possible to detect conventional fluorephores (FITC, PE, APC, GFP, RFP, mcherry) along with tandem dyes (PE-Cy5.5, PE-Cy7). Potential use includes detection of surface proteins for immunophenotyping, detection of transfected cells, identification of subpopulations based on size and granularity, detection of fluorescence properties of beads coated with nanoparticles.

Name of Instrument , manufacturing Company & Country: ​BD Fortessa X20 Flow cytometer , Becton Dickinson and Company (BD), USA
Laser Wavelength (nm) Comments
Red 637 Standard
Yellow-Green 561 Standard
Green 488 Standard

Applications 
a) Cellular uptake studies- Used for the quantitation of various fluorescently labelled drugs/ molecules into the mammalian as well as plant cells. It allows us to perform multiparametric analysis in the same set of cells with higher accuracy and reproducibility as compared to other techniques. 

b) Cancer Studies- Measures the number of cells in a sample, the percentage of live cells and dead cells can also be analyzed when treated with certain dyes such as propidium iodide (PI). The presence of tumor markers on the cell surface can also be detected when labelled with tumor specific fluorophore/ antibodies. 

c) Detection of apoptosis with Annexin V- Annexin V conjugates provide quick and reliable detection for studying the externalization of phosphatidylserine, an indicator of intermediate stages of apoptosis. The difference in fluorescence intensity between apoptotic and non-apoptotic cells stained with our fluorescent annexin V, as measured by flow cytometry, is typically about 100-fold. 

d) Cell Cycle analysis - Cellular DNA content can be analyzed following cell staining with 4',6'-diamidino-2-phenylindole (DAPI). This approach reveals distribution of cells in three major phases of the cycle (G1 vs S vs G2/M).

Samples required for FACS
Note: Samples state should be cell suspension, Cell pellet properly suspended in Phosphate buffer saline 1x (vol 100µl) and positive and negative controls are mandatory.  
Samples in suspension need to be given to the operator along with the following details: 
1. Type of sample/cells 
2. Fluorophores present 
3. Brief description of analysis required: For example, cell cycle analysis or percentage of bead bound to nanoparticles The operator will then run the samples and help with analysis on FACS DIVA software. Data will be exported and given as .fcs (raw data) and .pdf (analysed data) files. Additional analysis if required may be done by booking separate slots at nominal cost. • For cells prone to clumping: Cells should be suspended in 1X PBS with 0.1 mM EDTA (pH 8.0) and/or passed through a 70 µm nylon mesh before analysis.

Forbidden samples: Live bacteria will not be run on the flow cytometer. However, heat killed or aldehyde fixed bacteria may be run with prior permission. Similarly, contaminated cells should be avoided. • Data will be provided through google drive to the e-mail ID of the users. Users must collect the data on the day of the experiment. Data will be deleted on the 1st of every month
Testing Charges (Click here)

How to contact

Lab-Physical Location: 

First Floor, 
Kusuma School of Biological sciences IIT-Delhi, Hauz Khas, New Delhi – 110016 
Phone number : 011-2659-7503

Lab Coordinator

Prof. Anita Roy, 
Kusuma School of Biological Science  
IIT- Delhi, Hauz Khas, New Delhi – 110016 
Phone : 011 ​2659-7522
Email : ​anita.roy@bioschool.iitd.ac.in

Operational In-charge

Mr. Saurabh Shrivastva, 
Kusuma School of Biological sciences 
IIT-Delhi, Hauz Khas, New Delhi – 110016 
Phone number: ​011 ​2659-7512 
Email : saurabh.shrivastva@bioschool.iitd.ac.in

Get CRF Registration for 
External users 

1. Registration process (One Time): New Registration for external users.User Verification: Upload photo of Front side and back side of valid Identity card issued by the organization (only .jpeg format, file Size less than 1 Mb, height less than 1000 pixels, width less than 800 pixels). Undertaking: Download the undertaking template ( Download Link ). Duly filled, signed and stamped form should be uploaded in .pdf format. Confirmation: An email will be sent to the user after verification of the documents.

2.Before planning to avail the Instrument facility of CRF IIT Delhi: user must visit https://crf.iitd.ac.in -> Facility -> select the desired facility from left side navigation list.  Read carefully all the relevant information such as required sample type, testing charges and instrument location. Be prepared with details of the samples.

3.Log in and Click on Booking Appointment Tab: All the information relevant to testing must be filled in instrument booking form. In case user does not provide the required information the booking is liable to be rejected. User can upload the relevant existing data of their samples that might help in the measurements.

4.Sample submission & Payment Process: Once the application is submitted, the in-charge will verify the details and approve the request.External users must wait for the approval before making the payment and submitting the samples to concerned lab. After approval the payment should be made by the user. The payment transaction details need to be submitted online before appointment date time.

5.CRF Users can pay amount online using following details:
Beneficiary/Customer's Name: IRD ACCOUNT IITD
Bank Account Number:  10773572600  
Bank Name: State Bank of India Branch Name: IIT Branch
Branch Code: 01077
NEFT IFSC CODE / RTGS: SBIN0001077
please write "CRF _Instrument Name" in the Remarks section while paying. Save the receipt file and upload in the crfbooking system online against your application.Payment can also made in form of demand draft in favour of " IRD ACCOUNT IITD " Payable at New Delhi.6.IIT Delhi 
GST Number : 07AAATI0393L1ZI
PAN Number : AAATI0393L

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