Sandor LifeSciences
Biosciences Research Services

Flowcytometry


Flow cytometry department at Sandor life sciences provide the services of state of art multi-parameter cellular analysis for research and diagnostic samples. The facility currently houses the BD LSR II analyser equipped with 488 & 633 nm lasers and capable of detecting up to 7 different colours. The department provides training as well as support in designing an experiment and analysing the data to researchers and to the local scientific community. Please feel free to contact us for any queries and for any special need for your sample analysis.

Cell markers are an useful way to identify the cellular state and phenotype. Most of the immunological markers are CD (Cluster of differentiation) markers and are routinely used in immunological diagnostic and research studies. Our Immunophenotyping services enable you to identify and quantify your immune cells of interest. We can detect and quantify the surface, cytosolic and nuclear CD antigen expression on immune cells of your interest.

Surface CD Markers
Cytosolic CD markers
Nuclear CD markers

There is no single characteristic that adequately identify the stem cell by itself. We need to look at multiple cell surface markers and physiological characteristics. Human and rodent stem cell identification at Sandor life sciences is based on the expression of the listed surface markers. In addition to the listed surface markers we can custom design the panel according to your research interest.

Human Hematopoietic stem cell Markers

  • CD133
  • HLA-DR
  • CD7
  • CD34

Mouse Hematopoietic stem cell Markers

  • CD34
  • Sca-1
  • c-kit

Human Mesenchymal Stem cell

  • CD90+
  • CD44+
  • CD73+
  • CD45-ve
  • CD34-ve

Mouse Mesenchymal Stem cell

  • CD29 +
  • CD44 +
  • CD73 +
  • CD105+
  • CD106+
  • Sca-1
  • CD45-ve
  • CD11b-ve

Cell proliferation assays are performed to study the cell growth and differentiation. Proliferation assays are measured based on either the average DNA content or on cellular metabolic parameters. At Sandor life sciences we perform three different cell proliferation assays.

The determination of the cell cycle analysis is a well known technique in flow Cytometry

  • The cells stained with a nuclear staining dyes can be analysed by flow cytometry
  • Depending on the intensity of the stained dye we can proportionally estimate the quantity of DNA present in a cell.

CFSE labeling of live cells are useful for both invitro and in vivo studies and CFSE labeled cells can be used in tracking the survival of different cell populations. Flow Cytometry detection of up to eight successive cell divisions is possible. CFSE labeling can be coupled with other surface and intracellular markers if needed. We here at SANDOR is capable of detecting up to 7 different parameters of a dividing cell.

Flow cytometric analysis of BrdU incorporation assay determines the percentage of cells in S-phase that have incorporated a thymidine analogue, bromodeoxyuridine (BrdU), present in the culture medium during DNA replication. By coupling BrdU incorporation assay with a total DNA binding dye such as the PI, we can enumerate the cells that are actively synthesizing DNA with respect to their cell cycle positions.

An apoptosing cell has unique morphological and biochemical features. These unique hallmarks can be identified by flowcytometry in a cost effective and timely manner. At Sandor life sciences we can identify an individual apoptosing cell in three different methods

Activation of caspases are hallmarks of apoptosis and can be detected by Flowcytometry. Caspases play an important role in apoptosis and reagents that irreversibly bind to activated capases are tagged with fluorescent markers, to directly measure apoptosis as expressed by the number of active caspase enzymes present in the cell. Fluorescent Labelled Inhibitors of Caspases(FLICA), in combination with any DNA binding dye can be used to asses the cells in early apoptosis, late apoptosis, and necrosis stages.

An important early event in an apoptosing cell is the flipping of phosphatidylserine from the inside of the cell membrane to the out side. Annexin antibodies against Phosphatidylserine detects the presence of an early apoptosing event in a given cell. Simultaneous labeling with a DNA dye like Propidium Iodide enables us to precisely enumerate early apoptosing and late apoptosing cells using Flow Cytometry.

Proteins like H2AX and PARP play a significant role in DNA damage and repair. By flowcytometry we can detect the altered proteins and enumerate the number of cells undergoing DNA damage.

The occurrence of Micro nucleus inside the cell demonstrate high concordance with chromosomal aberrations. Screening for micronucleus is now a routine test in drug safety and toxicological studies. Flow cytometric detection of micronucleus is a cost effective and large samples can be tested in a less time consuming manner. DNA strand breaks are indicative signs of apoptotic cells and TUNEL assay can be detected by flow Cytometry in a less labor intensive manner.

Micro Nuclear assay
TUNNEL Assay
Intracellular Cytokine Assay
Cytokine Bead Assay

Errors occurring during mitosis and meiosis in plants can lead to multiplication of the entire genome. This phenomenon called pluriploidy can be assessed by flow Cytometry and the genome content can be calculated.

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