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Multiplexed Imaging of Gene Expression with CELESTA Light Engine

MERFISH (multiplex error robust fluorescence in
situ hybridization) is an imaging technique that
profiles cell populations based on the
identification of thousands of RNA transcripts per
cell. The CELESTA Light Engine is an ideal and widely-adopted illumination source for this
application. In a recent paper published in
Nature[1], Wheeler and co-workers used
MERFISH imaging with a CELESTA Light Engine
to quantify the expression of nine specific
astrocyte and T-cell markers. Five of the CELESTA Light Engine’s seven laser lines were used in the highly multiplexed MERFISH imaging protocol. The overall objective of the research described in the paper was to characterize astrocyte populations that contribute to pathogenesis in a preclinical model of multiple sclerosis.

Brain Organoid

50 micro thick slice cut from a brain organoid structure derived from induced pluripotent stem cells

Microscope: Nikon Ti2
Objective: Nikon 60X oil Plan Apo Lambda, NA 1.4
SD confocal module: Crest X-Light V3 
Confocal Laser bank: Lumencor CELESTA Light Engine

Staining: 
•	Magenta: TBR1 stained with Alexa 750. TBR1 is a transcription factor important for the differentiation of the nuclear glutamatergic neurons during cortical development. Staining specific for layer VI neurons.
•	Green: MAP2 stained with Alexa 488.  MAP2 is a protein associated with the microtubules present in the cytoskeleton of the neuronal dendrites.
•	Blue: DAPI