SSBD:database

421-Takenouchi-AgedYoungOocyte/Fig5_Chr1and18_20230204_1_Aged_5_lsm

Author reviewed

Time-lapse and z-series images of H2B-SNAP and the tdTomato-CENPC in aged mouse oocyte

SSBD:downloads

Project

421-Takenouchi-AgedYoungOocyte

Dataset ID

Fig5_Chr1and18_20230204_1_Aged_5_lsm

Kind

Image data

Title

Time-lapse and z-series images of H2B-SNAP and the tdTomato-CENPC in aged mouse oocyte

Description

Time-lapse and z-series images of H2B-SNAP and the kinetochore marker tdTomato-CENPC from prometaphase to anaphase of 16 to 22 month-old-mouse oocyte. The images of chromosome 1 and chromosome 18 at a at a certain time during imaging were reposited as DE_2_W000x_P001_T0yyy.lsm file.

Folder Name

Fig5_Chr1and18_20230204_1_Aged_5_lsm

File Format

.lsm

File Size

1.6GB

File Separation Method

Figure Number

Fig5

Dimensions

256x256x17x3x200

X Scale

0.119 micrometer

Y Scale

0.119 micrometer

Z Scale

1.5 micrometer

T Scale

3 minutes

Channel

3 channel

Quantitative Data

Paper(s)

Biosample

Description

From prometaphase to anaphase oocyte from 16 to 22 month-old-mouse, in which chromosomes and kinetochores were visualized.

Organism

Mus musculus (NCBITaxon:10090)

Strain

B6D2F1/J (https://www.jax.org/strain/100006)

Cell

oocyte (CL:0000023)

Cell line

MeSH

Oocytes (D009865)

Microscopy, Confocal (D018613)

Meiosis (D008540)

Chromosomes (D002875)

Chromosome Segregation (D020090)

Intrinsic Variable

Extrinsic Variable

To visualize chromosomes and kinetochores, in vitro transcribed mRNAs for H2B-SNAP and tdTomato-CENPC (centromere protein C) were injected. To detect chromosome1 and chromosome18, MCP-3tagBFP and PCP-3mClover were also introduced for CRISPR-Sirius method.

Ontology

Anatomical Entity

UBERON

oocyte (CL:0000023)

Biological Process

female meiosis chromosome segregation, (GO:0016321)

Cellular Component

chromosome (GO:0005694)

Molecular Function

MeSH Anatomy

Oocytes (D009865)

Microscopy, Confocal (D018613)

Meiosis (D008540)

Chromosomes (D002875)

Chromosome Segregation (D020090)

MeSH BP

Oocytes (D009865)

Microscopy, Confocal (D018613)

Meiosis (D008540)

Chromosomes (D002875)

Chromosome Segregation (D020090)

MeSH CC

Oocytes (D009865)

Microscopy, Confocal (D018613)

Meiosis (D008540)

Chromosomes (D002875)

Chromosome Segregation (D020090)

MeSH MF

Oocytes (D009865)

Microscopy, Confocal (D018613)

Meiosis (D008540)

Chromosomes (D002875)

Chromosome Segregation (D020090)

Genetics

Gene

Protein / Probe

Histone H2B type 1-J

Method / Genotype

injection

Gene

Protein / Probe

Centromere protein C

Method / Genotype

injection

Gene

Protein / Probe

-single-guide RNA targeting repeat sequences of the chromosome1

Method / Genotype

Gene

Protein / Probe

-single-guide RNA targeting repeat sequences of the chromosome18

Method / Genotype

injection

Treatments

Reagent / Compound

Concentration

Fold Dilution

Imaging Method

confocal microscopy (FBbi:00000251)

time lapse microscopy (FBbi:00000249)

Detection Method

photomultiplier tube (PMT)

Visualization Method

H2B with SNAP-tag was visuallised with SNAP-Cell 647-SiR. To visualize kinetochores, centromere protein C was tagged with tdTomato. Chromosome1 and Chromosome18 were visualized with BFP and mClover respectively.

Illumination Method

single point scanning

Source of Contrast

distribution of a specific protein

Imaged Parameter

fluorescence emission

Sample Preparation Method

living tissue

MeSH

Oocytes (D009865)

Microscopy, Confocal (D018613)

Meiosis (D008540)

Chromosomes (D002875)

Chromosome Segregation (D020090)

Instrument

Body

Zeiss LSM780

Module

Light Source

Detector

Objective

Zeiss 40×C-Apochromat 1.2NA

Filter Set

Dichroic