title: GSM8759172: Ovary slow-frozen and vitrified, slice 1 & 2, 28 yo donor; Homo sapiens; OTHER
library_name: GSM8759172
library_construction_protocol: Human ovarian tissues were donated from one patient aged 35 years who underwent oophorectomy due to cervical cancer. The ovarian tissues were removed using scissors rather than electrocoagulation. Obtained ovarian tissues were directly placed in sterile transport medium (Leibovitz抯 L-15 Medium supplemented with 1% HSA) and transported on ice to the laboratory. In the laboratory, the ovarian tissues were transferred to 10 cm sterile dishes containing chilled sterile normal saline. The saline was changed several times during the process. After most of the medulla was removed with scalpels, the tissues were cut into 10?? mm pieces and divided into fresh and two cryopreservation groups. According to the Stereo-Seq protocol, one piece of fresh group tissues was fixed in O.C.T. compound (Tissue Tek) for subsequent sequencing, tissues of the two cryopreservation groups were processed after freezing and thawing. The tissue capture area was approximately 10? mm. 10?m tissue sections were cut using cryo-microtome (CM1950, Leica) with head temperature set at -16 癈 and adhered to the Stereo-seq chip (BGI) surface. The sections were then performed histomorphology detection, staining in hematoxylin for 3 min, 10 s in bluing agent, and 1 min in eosin. To release mRNA from the cells, the sections were permeabilized, the optimal time was 22 min. After in situ reverse transcription, the resulting cDNAs were amplified. PCR products were purified for DNA nanoball (DNB) generation and finally sequenced on MGI DNBSEQ-Tx sequencer; detailed protocols are available on the BGI Stereo-seq website (https://en.stomics.tech/).