Microbiology and Biotechniques

Question: Discuss about theMicrobiology and Biotechniques. Answer: Review of the Literature Flow Cytometry The technique or the special type of technology that is used for analyzing the chemical and the physical characteristics of any particles present in a fluid that are allowed to passed from a laser technique, that are labeled and then are excited by the laser process by emitting light at varying at different wavelength (Givan 2013). The determination of the fluorescence has various different properties of single particles. The properties are measured in aspect to the relative granularity, sizes and the intensity as well as the complexity of the internal part of the particle. The Lasik optic system illuminates the particles that are present in the stream for passing and scattering of the light. Any particle that illuminated the fluorescent light is captured by the carefully positioned lens. The light scatters at two different angles. The data is mainly collected from each of the particle that is determined on the basis of the scattering of the light and the fluorescent properties (Piyasenaand Graves 2014). The data of the cytometry flow is presented in the form of histograms or as in the form of the parameters that is correlated, referred as cytograms. Several Researches have taken place based on the flow cytometry but analyzing of the micro biota that is associated with the midguts of the vector mosquito marks as an important milestone on the research of the flow cytometry (Habtewold, Duchateau and Christophides, 2016). Understanding the structure and the function of the microbiota that is related with the midgut of the mosquitoes as the vector causing disease is increasing day by day. The growing phase of this knowledge has faced several limitations and the challenges that is related with the modern culture and the PCR techniques. Flow cytometry can be combined with different cell dyes that are being applied in ecological microbiology. It is also being coupled with dead or living in the staining procedure with the dyes. Flow cytometry also helps in the differential dyes that are used for the staining purposes. It also shows a close bonding between the corresponding dilute factors along with the cell counts and this technique will have better precision that is compared to qRTPCR. The FCM count the microbiota of the mosquito reaches to the peak load after 18hours of post feeding and reduces approx after 24 hours (Habtewold, Duchateau and Christophides, 2016). The midgut of the mosquito is the main targets for controlling of the diseases, the study that focuses on the interactions with the vectors of the mosquito and the pathogens received a great attraction. An aspect of these investigations involving the establishments of the impact of the immune system of mosquito along with the genes that is involved in establishment of the genes that is related to the immune genes using a special process called RNAi based gene slicing technique where homologous mRNA becomes the target gene that destroys by the action of dicer machinery. The bacteria in the gut of the mosquitoes cannot be cultivatable under any special conditions and cannot be observed using the culture dependant techniques. This limitation is being removed by the development based on the cultures that is independent in techniques. Flow cytometry can be combined with different techniques of the cell training and is successful in applying in the field of the ecological microbiology. Fluorescent labeled antibodies are used for marking the bacterial cells, general DNA binding dyes like SYBR green and PI for the analysis of Flow cytometry. Machines that are used for FCM are FACS Calibur flow cytometer, mainly for the proof of direct analysis of the microbiota present in the midgut. The other analyzers that are used for the analyzing and optimizing the fluorescent signals called BD LSRFortessa. These are used for evaluating and optimizing the fluorescent signals on SYBR and PI channels. FACSAria is another type of FCM that is used for sorting down the cells (Habtewold, Duchateau and Christophides, 2016). Analyses in conjunction with SGI/PI staining can forcefully discriminate between the dead and the living or dead microbial cells and can separate both the types from auto fluorescing debris in a high, dependable and in very precise fashion. This can be applied directly on the midgut homogenates and offers an inexpensive option for qualifying the microbiota as it may passes down the intermediate steps. FCM also depends on the availability of species-specific antibody and can be used for the detection and the quantification of the gut of the microbiota to the species that can be achieved. The strengths of Cytometry flow (Herculano-Houzel et al. 2015): Helps in measuring single cells Multiple analyses of the parameters Small populations are being identified Predefined cell population were sorted The weaknesses are as follows (Herculano-Houzel et al. 2015): Sophisticated and expensive instruments Single cell particle requires Structure of the tissue gets lost Flow cytometry are used for analyses for the evaluation of the trueness for the somatic embryogenesis that regenerates the plants tissue by tissue culture method. It is mainly done to check the ploidy of the somaclonal variation or by the separation technique that is genotypically different from the other cell layers. Flow cytometry was used mainly for the programs related to breeding techniques. RQ- Can flow cytometry revolutionize the conventional methods of tissue culture? References List: Givan, A.L., 2013. Flow cytometry: first principles. John Wiley Sons. Habtewold, T., Duchateau, L. and Christophides, G. (2016). Flow cytometry analysis of the microbiota associated with the midguts of vector mosquitoes. Parasites Vectors, 9(1). Herculano-Houzel, S., von Bartheld, C.S., Miller, D.J. and Kaas, J.H., 2015. How to count cells: the advantages and disadvantages of the isotropic fractionator compared with stereology. Cell and tissue research, 360(1), pp.29-42. Piyasena, M.E. and Graves, S.W., 2014. The intersection of flow cytometry with microfluidics and microfabrication. Lab on a Chip, 14(6), pp.1044-1059.