BIOMAG

Image Source: BD Biosciences, BD Influx Cell Sorter Waste Tank : >Disconnect the Vacuum Pump and Fluidic Lines. >Open Waste Tank and Add 1% Bleach and discard. >Close the lid tightly (To ensure vacuum build up). >Reconnect the vacuum Pump and Fluidics Lines. Sheath Tank : >Release pressure using pressure knob. >Disconnect all the fluid lines connected to the sheath Tank. >Open and check the sheath level (if required, add sheath fluid to the sheath tank). >Close and connect the air and fluid lines. > Connect Sheath Line to Filter to Instrument. Switching ON : >Switch on Condenser, Vacuum Pump, Mains of the Instrument and Computer. >Wait for around 10 minutes to build the pressure in the waste tank and sheath tank. >Wait for 10 minutes for lasers to fully warm up and turn them ON one by one. >Make Sure, the displayed sheath pressure matches the pressure build scale on the sheath tank. >Attach the nozzle to the nozzle asse

Fluorescence Microscopy and Flow Cytometry, both are biological instruments which are capable of measuring the biophysical and biochemical properties of cells or particles which are under study. Both of them are capable of identifying types of cells present in a given sample. These two technologies are useful in variety of applications and they have widespread usage... But, These two differ in their applications and use.  Fluorescence Microscopy Flow Cytometry Sample mounted on a glass slide. Sample is allowed to pass through a liquid stream through a sensing area. (Single cell suspension). Produces magnified images of the cells Does not produce the images. Whole Tissue section can be visualized and analyzed Tissue must be disintegrated into single cell suspension. And each single cell is analyzed. Detection through human eye and images can be produced on a computer screen Detection is e

Please Watch The Demo Video attached after the protocol and example Data!!!! Preparation of Samples for Cell Cycle using Flow Cytometry :  1) Suspend approximately million cells (10^6) in 1mL of 1x PBS (phosphate-buffered saline solution). and with the help of pipette, gently aspirate (up and down) about 7-8 times, to make the single cell suspension.  2) Take a centrifuge tube and add 4mL of 70% ethanol.. (70% ethanol should be in ice before using). and add 1mL of cells in PBS to the same tube. Keep the tubes on ice for minimum 2 hours. This step is to fix the cells. (Cells can be kept at 4℃ for a week). 3) Centrifuge the ethanol fixed cells for 5 minutes at 300g. Carefully take out the supernatant. Do not disturb the pellet.  4) Add 5mL of cold 1xPBS to this pellet and suspend the cells using pipette.  5) Centrifuge the tubes at 300g for 5 minutes. Discard the supernatant. Again do not disturb the pellet.  6) If required, step 5 can be repeated f

Biotech and Med Information Hello Everyone,  Please Join Our Facebook page.

Immunophenotyping is a most important and regularly used technique  in Immunology and Immunological research studies. The technique is usually used for the disease monitoring and diagnostic purpose either by immunohistochemistry or by utilizing the high-throughput technique Flow Cytometry.. Flow Cytometry is a common method for the immuno phenotyping the different cell subsets or sub populations of cells. Immunophenotyping is used for the identification and characterisation of the cell surface markers and intracellular markers. In simple terms, the technique is used to detect and study the proteins expressed by the cells. Samples used for the study could be Tissue Section (Fresh or Fixed) or Cell suspension (mono-dispersed cell suspension for FCM). This technique helps in rapid and easy phenotyping of each cell in a heterogeneous sample according to the presence or absence of a protein combination. Markers (or proteins of interest) are usually functional membrane proteins whi

Flow Cytometry Analysis using Peripheral Blood Cells: 1) Collect approximately 75 microliters of blood into 1mL PBS containing 5microM EDTA and mix immediately to prevent clotting. keep the tubes on ice. (10 microliters of 0.5M EDTA stock). 2) Remove Red blood cells from samples. [RBCs are lysed using either Gey's solution or a buffered ammonium chloride solution. (Or, BD sells a product called FACS lysis buffer" that is used after the staining protocol to lyse RBCs and fix the cells)]. Use proper solution according to the instructions given by the manufacturer.  3) Wash the cells at least 2-3 times with FACS buffer. (FACS buffer contains PBS solution supplemented with either 1% BSA or 5% FBS and 0.05% NaN3).  4) Suspend the pellet from the final wash in 50microliters FACS buffer per each analysis on a single sample. (Up to three separate staining reactions can be set up from a single sample).   5) Add 50 microliters of cell suspension to 10 microliters of