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Sheba Medical Center

3 ARTICLES PUBLISHED IN JoVE

Medicine

Cell Death Associated with Abnormal Mitosis Observed by Confocal Imaging in Live Cancer Cells

Asher Castiel¹, Leonid Visochek^2,3, Leonid Mittelman⁴, Yael Zilberstein⁴, Francoise Dantzer⁵, Shai Izraeli^1,6, Malka Cohen-Armon^2,3

¹Cancer Research Center, Sheba Medical Center, ²The Neufeld Cardiac Research Institute, Tel-Aviv University, ³Department of Physiology and Pharmacology, Tel-Aviv University, ⁴Imaging Unit, Sackler Faculty of Medicine, Tel-Aviv University, ⁵Biotechnology and Cell Signaling, Ecole Superieure de Biotechnologie Strasbourg, ⁶Department of Human Molecular Genetics and Biochemistry, Tel-Aviv University

The cytotoxic activity of the phenanthridine PJ-34 in cancer cells undergoing mitosis was documented in real time by live confocal imaging. PJ-34 eradicated human breast cancer MDA-MB-231 cells harboring extra-centrosomes in mitosis. Unlike normal bi-focal mitosis, the extra-centrosomes were not clustered in the two spindle poles in the presence of PJ-34.

Biology

Guided Protocol for Fecal Microbial Characterization by 16S rRNA-Amplicon Sequencing

Ayelet Di Segni¹, Tzipi Braun¹, Marina BenShoshan¹, Sarit Farage Barhom¹, Efrat Glick Saar¹, Karen Cesarkas¹, James E. Squires², Nathan Keller^1,3, Yael Haberman^1,4

¹Sheba Medical Center, ²Children's Hospital of Pittsburgh of UPMC, ³Ariel University, ⁴Cincinnati Children's Hospital Medical Center

This manuscript describes a detailed standardized protocol of high-throughput 16S rRNA-amplicon sequencing. The protocol introduces an integrated, uniformed, feasible, and inexpensive protocol starting from fecal sample collection through data analyses. This protocol enables analysis of large numbers of samples with rigorous standards and several controls.

JoVE Core

T and B Cell Receptor Immune Repertoire Analysis using Next-generation Sequencing

Lael Werner^1,2, Chen Dor^3,4, Naomi Salamon¹, Meital Nagar^3,4, Dror S. Shouval^1,2

¹Pediatric Gastroenterology Unit, Edmond and Lily Safra Children's Hospital, Sheba Medical Center, ²Sackler Faculty of Medicine, Tel Aviv University, ³Cancer Research Center, Sheba Medical Center, ⁴Molecular Hematology Laboratory, Sheba Medical Center

The current protocol describes a method for DNA isolation from blood samples and intestinal biopsies, generation of TCRβ and IGH PCR libraries for next-generation sequencing, performance of a NGS run and basic data analysis.