Is it beneficial for a geneticist to have theoretical and practical courses on flow cytometry the software using and Its a kind of cell sorting and counting thing I guess so for a geneticist is it important to what extent?Does it have any applications in the stem cell and regeneration technology?
Flow cytometry and cell sorting is quite widely used in stem cell research, at least if you study haematopoietic stem cells. It is definitely worth knowing the basics so you can understand research papers.
As for actual practical experience, it depends on what you want to research. Cell sorting is commonly used as a way of isolating the cells you want, at the right developmental stage etc so you can further analyse them, for example using a more molecular biology approach.
Discuss with your supervisor/line manager whether using flow cytometry would be useful to answer your particular research question. Maybe, you can just learn the theory and collaborate with another group that have the experience? Or if it is easier, get the experience by learning from someone who uses flow cytometry a lot.
I haven't dealt with it, and not even my faculty yet, and I want to attend a course on it and it will cost me at least 1000 dollars for about 15 days, and in my PhD I look forward to have my research on stem cells and these isolating cells so I want to know in a sense that to what extent it can help me to understand ll about it and use of it
Absolutely, "YES".
With the increased sensitivity of genetic analyses, the single cell analyses are becoming more and more important. The single cell analyses by FACS will give you the information of gene mutations and expression in each type of cells.
It is not so much important when you are just seeking for causative mutations in certain genetic disorders. But, still you may need to examine gene expression profiles in cells affected in the diseases, which will definitely need cell sorting.
In the field of oncology and stem cell biology, it is needless to say. You must know how to sort stem cells, differentiated normal cells, cancer cells, cancer stem cells, cancer-associated fibroblasts, etc. It is surely enhance the quality of your research.
Regards,
In the broader sense the answer to your question is yes. The cell sorting, counting, propagation so on used in different areas of research not only in molecular biology but also in cancer research, and various kind of therapeutic, drug and diagnosis research.
There are very sophisticated and manual methods as well, for ell sorting and counting, The method selection is depend on your research topic. Because the purity of sorted cells you need depends on the down stream application of sorted cells.
First, you go carefully through few full text articles on the related topic, and then you may get a better idea to discuss it further with eminent researches.
You can check this:
http://isac-net.org/News/Society/How-is-Cytometry-the-Driving-Force-in-Stem-Cell-Re.aspx
Characterization of stem cell cultures falls into two main categories: first, monitoring the genomic integrity of the cells and second, tracking the expression of proteins associated with pluripotency. Genomic analysis is necessary to ensure stem cells maintained in culture for long periods of time have not become unstable through chromosomal loss or duplication, or changes in their epigenetic profiles. Proteomic analysis ensures that the cells are expressing the factors necessary to maintain pluripotency. Finally, analysis of the differentiated state by analyzing key genetic and protein markers will ensure identification and propagation of the proper cell type This section provides an overview of different analysis methods for stem cells such as flow cytometry.
Flow cytometry with cell sorting is a method for sorting a heterogeneous population of cells based on their light scattering characteristics,. Fluorescently tagged monoclonal antibodies are used to distinguish a specific cell population. Stem cells express a number of cell surface proteins that are considered to be markers of pluripotency and many antibodies are available for their detection. Glycolipid antigens SSEA-3 and SSEA-4 and the keratin sulfate antigens TRA-1-60 and TRA-1-81 are commonly used antigens for identifying and sorting live stem cell populations. Fixed stem cells can also be sorted based on the expression of the transcription factors Oct-3/4 and NANOG. As stem cells differentiate, the loss of these markers and expression of new markers can be used to track the progress and overall condition of a stem cell population. For instance, Nestin is a protein marker for neural progenitor cells, BNP (brain natriuretic peptide) is a marker for cardiomyocytes, and CD31, CD34, and CD45 are markers for hematopoietic cell types.
Genetic application
http://www.medscape.org/viewarticle/571611
Check out the special issue of Cytometry Part A from january 2013, Volume 83A, Issue 1, Pages 1–169, for a taster of the potential of flow cytometry. Simple evaluation of ES status via SSEA ± TRA surface. Assessment of lineage differentiation through markers of the cell surface, cytoplasm and nucleus. Ultimately, allowing identification of subpopulations of interest for sorting for subsequent molecular analysis .
Absolutely, "YES"
Flow cytometry, cell sorting, cell counting absolutely important in the field of genetics and stem cell research.
This is the basic experiment you have to do when you work with stem cell biology. I guess you want to know how pure your cell population.
I am interested in studying toxicity of different nanomaterial on live cells. Using scattering and fluorescence detection simultaneous, how can I see difference in the behavior of live cells or cells having certain dozes of certain nanoparticle inside the cells. What do you suggest the material which I should start with and what kind of live cells? Does live cells absorb nanoparticle in the cells without any hindrance from the cell membrane? If any one has deeper study , please write me or refer me a material for it. I will appreciate for your help.
Cell sorting is particularly important for RNASeq and methylation analysis (and other real-time genetic analysis techniques) when you have a hypothesis that depends on a particular cell type. Sorting reduces the variation that is due to different cells having a different genetic structure or expression.
Yes, it is. For example if you want to analyse gene expression pattern in the peripheral blood due to some disease or pathogen challenges you never know which immune cell type react in which way if you wouldn't apply a cell sorting before.
The techniques are easy to learn however they are a bit costly. It would be fine , if you have access to a flow cytometer or a device for magnetic cell separation.
Best regards
Dirk
- Badges
- Science method