Bushong, Eric A.; Martone, Maryann E.; Ellisman, Mark H. (2021). CIL:36304, Rattus, astrocyte. In Cell Image Library. UC San Diego Library Digital Collections. Dataset. https://doi.org/10.6075/J0NK3F2M

Intracellular dye injections of protoplasmic astrocytes from the CA1 region of hippocampus of 3 week old rat reveals highly ramified spongiform processes that span territories with minimal overlap. Astrocytes were filled with Lucifer Yellow (green) and Alexa 568 (red) Optical sections were generated with a single photon confocal microscope.

• 2021

• Bushong, Eric A.
• Ellisman, Mark H.
• Martone, Maryann E.

Male Sprague Dawley rats, 3 weeks of age, were anaesthetized and perfused transcardially with with Ringer's solution (37°C), followed by 4% paraformaldehyde (PFA) in PBS (37°C, pH 7.4). Tissue was sectioned (100 µm thick) using a vibratome, and injected with a BX50WI microscope (Olympus, Melville, NY) and, using infrared differential interference contrast videomicroscopy,cells with a small diameter soma (approx. 7–11µm) in the stratum radiatum of CA1 were located and impaled with glass micropipettes filled with either 5% aq. LY, 20mM AlexaFluor 568 in 200mM KCl, or 20mM AlexaFluor 488 in 200mM KCl. The astrocytes were filled using a 0.5 Hz pulses of current until all of the processes were brightly fluorescent. After several astrocytes were filled, a slice was placed in the 4% PFA overnight at 4°C. Slices were then imaged after coverslipping in Gelvatol. Confocal images were acquired with a BioRad Radiance2000 (Hercules, CA) attached to a Nikon E600FN (Kanagawa, Japan) microscope equipped with a Nikon (Tokyo, Japan) 60× oil objective (NA 1.4). Optical resolution of the z-axis, 0.25µm. Additional details are provided in Bushong EA, Martone ME, Ellisman MH. Maturation of astrocyte morphology and the establishment of astrocyte domains during postnatal hippocampal development. Int J Dev Neurosci. 2004 Apr;22(2):73-86.

Preparation: formaldehyde fixed tissue
Relation to intact cell: vibratome-sectioned tissue
Item type: recorded image
Imaging mode: confocal microscopy
Parameter imaged: fluorescence emission
Source of contrast: intracellular filling
Visualization methods: lucifer yellow; Alexa Fluor 568
Data qualification: Processed;spatialmeasurements

• Cell Image Library Group ID: 10473

• Rattus

• Astrocyte
• Cell body
• Cell projection cytoplasm

• Forebrain astrocyte development

• data
• image

• No linguistic content; Not applicable

Samplenumber: 36304

Doi: https://doi.org/10.6075/J0NK3F2M

Source data

• Source Record in the Cell Image Library: https://doi.org/10.7295/W9CIL36304

Previous version

• Item in Cell Centered Database, UC San Diego Library Digital Collections: https://doi.org/10.6075/J09P31FH

Reference

• Abiodun MO, Johnson TO. Liver function tests in Nigerian diabetics with particular reference to brom-sulphalein retention. West Afr Med J Niger Pract. 1972 Apr;21(2):47-50. PMID: 5036382. https://www.ncbi.nlm.nih.gov/pubmed/?term=5036382

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2022-08-12