Confocal microscopy offers several advantages over conventional optical microscopy, including shallow depth of field, elimination of out-of-focus glare, and the ability to collect serial optical sections from thick specimens. In the biomedical sciences, a major application of confocal microscopy involves imaging either fixed or living cells and tissues that have usually been labeled with one or more fluorescent probes.
The human microbe ome is composed of communities of bacte-ria, viruses and fungi that have a greater complexity than the human genome it-self. Genome sequencing technologies and metagenomic analysis has helped in our understanding of human micro-biome. This is useful in manipulation of gut microbiome to be used in the treat-ment of childhood diseases.
Confocal microscopy, most frequently confocal laser scanning microscopy (CLSM) or laser confocal scanning microscopy (LCSM), is an optical imaging technique for increasing optical resolution and contrast of a micrograph by means of using a spatial pinhole to block out-of-focus light in image formation. Capturing multiple two-dimensional images at different depths in a sample enables the reconstruction of three-dimensional structures (a process known as optical sectioning) within an object. This technique is used extensively in the scientific and industrial communities and typical applications are in life sciences, semiconductor inspection and materials science
Light travels through the sample under a conventional microscope as far into the specimen as it can penetrate, while a confocal microscope only focuses a smaller beam of light at one narrow depth level at a time. The CLSM achieves a controlled and highly limited depth of focus.
DNA Sequencing System
Sequencing means finding the order of nucleotides on a piece of DNA. Nucleotide order determines amino acid order, and by extension, protein structure and function (proteomics). An alteration in a DNA sequence can lead to an altered or non functional protein, and hence to a genetic disorder. DNA sequence is important to detect the type of mutations in genetic diseases and offer hope for the eventual development of treatment DNA.
Methods of sequencing
1. Sanger dideoxy (primer extension/ chain-termination) method
Most popular protocol for sequencing, very adaptable, scalable to large sequencing projects.
2. Maxam-Gilbert chemical cleavage method
DNA is labelled and then chemically cleaved in a sequence dependent manner. This method is not easily scaled and is rather tedious.
It provides an important tool for determining the thousands of nucleotide variations associated with specific genetic diseases, like Huntington’s, which may help to better understand these diseases and advance treatment.