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#chromosome analysis

2 public questions tagged with this topic.

Plant tissue culture 3

Variant detection by cytological studies involves:

Cytological analysis visualizes nuclear integrity to uncover ploidy changes and structural rearrangements. Actively growing root tips or callus are pretreated with mitotic inhibitors such as colchicine or paradichlorobenzene to accumulate metaphases, fixed in Carnoy's fluid, hydrolyzed with HCl, and stained with acetocarmine, Feulgen reagent, or fluorescent DAPI binding AT-rich DNA. Squash preparations allow chromosome counting, measurement of arm ratios, and identification of deletions, duplications, translocations, and fragments under light or epifluorescence microscope. Monitoring lagging c

Ref: Sharma & Sharma Chromosome Techniques Butterworth; NCBI NBK21134 karyotyping; PLOS ONE garlic somaclonal cytology 2020; Lodish Ch 20 microscopy.

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Transcriptome analysis

What kind of microscope is used in FISH?

FISH signals are inherently fluorescent, generated by fluorophores such as FITC, TRITC, Texas Red, Cy3, and Cy5 attached to probes or detected via fluorescent antibody sandwiches. These faint nuclear signals require excitation with specific wavelengths and emission filter sets, which only fluorescence microscopy provides. Confocal microscopy further improves signal-to-noise ratio and optical sectioning, crucial for interphase nuclei and three-dimensional tissue FISH. Bright-field, electron, and phase-contrast microscopes cannot excite or separate fluorochrome emission, thus lacking capability

Ref: NCERT Biology Class XII Principles on Klenow fill-in labeling, Lehninger Chapter 9 DNA cloning techniques, and Molecular Cloning by Sambrook Chapter 10 documenting end-labeling of cohesive termini.

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