PMID- 35103978
OWN - NLM
STAT- MEDLINE
DCOM- 20220331
LR  - 20220422
IS  - 1940-6029 (Electronic)
IS  - 1064-3745 (Linking)
VI  - 2458
DP  - 2022
TI  - Superresolution Microscopy for Visualization of Physical Contacts Between 
      Chromosomes at Nanoscale Resolution.
PG  - 359-375
LID - 10.1007/978-1-0716-2140-0_20 [doi]
AB  - This protocol describes the fluorescence in situ hybridization (FISH) of DNA 
      probes on mitotic chromosome spreads optimized for two super-resolution 
      microscopy approaches-structured illumination microscopy (SIM) and stimulated 
      emission depletion (STED). It is based on traditional DNA FISH methods that can 
      be combined with immunofluorescence labeling (Immuno-FISH). This technique 
      previously allowed us to visualize ribosomal DNA linkages between human 
      acrocentric chromosomes and provided information about the activity status of 
      linked rDNA loci. Compared to the conventional wide-field and confocal 
      microscopy, the quality of SIM and STED data depends a lot more on the optimal 
      specimen preparation, choice of fluorophores, and quality of the fluorescent 
      labeling. This protocol highlights details that make specimens suitable for 
      super-resolution microscopy and tips for good imaging practices.
CI  - (c) 2022. The Author(s), under exclusive license to Springer Science+Business 
      Media, LLC, part of Springer Nature.
FAU - Yu, Zulin
AU  - Yu Z
AD  - Stowers Institute for Medical Research, Kansas City, MO, USA.
FAU - Potapova, Tamara A
AU  - Potapova TA
AD  - Stowers Institute for Medical Research, Kansas City, MO, USA. tpo@stowers.org.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Methods Mol Biol
JT  - Methods in molecular biology (Clifton, N.J.)
JID - 9214969
RN  - 0 (Fluorescent Dyes)
SB  - IM
MH  - *Chromosomes
MH  - *Fluorescent Dyes
MH  - In Situ Hybridization, Fluorescence/methods
MH  - Microscopy, Confocal/methods
MH  - Microscopy, Fluorescence/methods
OTO - NOTNLM
OT  - Chromosome spreads
OT  - Fluorescence in situ hybridization (FISH)
OT  - Immuno-FISH
OT  - Ribosomal DNA (rDNA)
OT  - Stimulated emission depletion (STED)
OT  - Structured illumination microscopy (SIM)
EDAT- 2022/02/02 06:00
MHDA- 2022/04/01 06:00
CRDT- 2022/02/01 12:15
PHST- 2022/02/01 12:15 [entrez]
PHST- 2022/02/02 06:00 [pubmed]
PHST- 2022/04/01 06:00 [medline]
AID - 10.1007/978-1-0716-2140-0_20 [doi]
PST - ppublish
SO  - Methods Mol Biol. 2022;2458:359-375. doi: 10.1007/978-1-0716-2140-0_20.