Microwave-Assisted B Cell Tomography

In vitro cultured B cells were fixed in Karnovsky’s fixative (Electron Microscopy Sciences) and processed according to the previously published microwave irradiation strategy^{56 (link)} with the following modifications: Cells were centrifuged at 800g between steps, cells were infiltrated in Araldite resin (SPI, Inc.) and tomography sections were cut at 150 nm thickness and imaged with no post-section staining. An Ultrascan 4000 camera (Gatan, Inc.) with SerialEM acquisition and control software (University of Colorado, Boulder, CO)^{57 (link)} was used to collect the tomographic tilt series.

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Akkaya M., Traba J., Roesler A.S., Miozzo P., Akkaya B., Theall B.P., Sohn H., Pena M., Smelkinson M., Kabat J., Dahlstrom E., Dorward D.W., Skinner J., Sack M.N, & Pierce S.K. (2018). Second signals rescue B cells from activation-induced mitochondrial dysfunction and death. Nature immunology, 19(8), 871-884.

Publication 2018

Karnovsky’s fixative Ultrascan 4000 camera

Araldite B cells Cells Electron microscopy Fixative Microwave irradiation Resin Tomographic

Corresponding Organization : National Institutes of Health

Other organizations : National Heart Lung and Blood Institute

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Variable analysis

independent variables

Microwave irradiation strategy

dependent variables

Ultrastructural characteristics of in vitro cultured B cells

control variables

Karnovsky's fixative
Araldite resin
Tomography section thickness
No post-section staining
Ultrascan 4000 camera
SerialEM acquisition and control software

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

Annotations

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