PMID- 32704503
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20240329
IS  - 2306-5729 (Print)
IS  - 2306-5729 (Electronic)
IS  - 2306-5729 (Linking)
VI  - 4
IP  - 3
DP  - 2019 Sep
TI  - TIRF Microscope Image Sequences of Fluorescent IgE-FcepsilonRI Receptor Complexes 
      inside a FcepsilonRI-Centric Synapse in RBL-2H3 Cells.
LID - 111 [pii]
LID - 10.3390/data4030111 [doi]
AB  - Total internal reflection fluorescence (TIRF) microscope image sequences are 
      commonly used to study receptors in live cells. The dataset presented herein 
      facilitates the study of the IgE-FcepsilonRI receptor signaling complex (IgE-RC) in rat 
      basophilic leukemia (RBL-2H3) cells coming into contact with a supported lipid 
      bilayer with 25 mol% N-dinitrophenyl-aminocaproyl phosphatidylethanolamine, 
      modeling an immunological synapse. TIRF microscopy was used to image IgE-RCs 
      within this FcepsilonRI-centric synapse by loading RBL-2H3 cells with fluorescent 
      anti-dinitrophenyl (anti-DNP) immunoglobulin E (IgE) in suspension for 24 h. 
      Fluorescent anti-DNP IgE (IgE(488)) concentrations of this suspension increased 
      from 10% to 100% and corresponding non-fluorescent anti-DNP IgE concentrations 
      decreased from 90% to 0%. After the removal of unbound anti-DNP IgE, multiple 
      image sequences were taken for each of these ten conditions. Prior to imaging, 
      anti-DNP IgE-primed RBL-2H3 cells were either kept for a few minutes, for about 
      30 min, or for about one hour in Hanks buffer. The dataset contains 482 RBL-2H3 
      model synapse image stacks, dark images to correct for background intensity, and 
      TIRF illumination profile images to correct for non-uniform TIRF illumination. 
      After background subtraction, non-uniform illumination correction, and conversion 
      of pixel units from analog-to-digital units to photo electrons, the average pixel 
      intensity was calculated. The average pixel intensity within FcepsilonRI-centric 
      synapses for all three Hanks buffer conditions increased linearly at a rate of 
      0.42 +/- 0.02 photo electrons per pixel per % IgE(488) in suspension. RBL-2H3 cell 
      degranulation was tested by detecting beta-hexosaminidase activity. Prolonged 
      RBL-2H3 cell exposure to Hanks buffer inhibited exocytosis in RBL-2H3 cells.
FAU - Drawbond, Rachel
AU  - Drawbond R
AD  - UCCS Center of the Biofrontiers Institute, University of Colorado at Colorado 
      Springs, Colorado Springs, CO 80918, USA.
AD  - Department of Mathematics, University of Colorado at Colorado Springs, Colorado 
      Springs, CO 80918, USA.
FAU - Spendier, Kathrin
AU  - Spendier K
AUID- ORCID: 0000-0002-5784-5253
AD  - Department of Mathematics, University of Colorado at Colorado Springs, Colorado 
      Springs, CO 80918, USA.
AD  - Department of Physics and Energy Science, University of Colorado at Colorado 
      Springs, Colorado Springs, CO 80918, USA.
LA  - eng
GR  - R15 GM128166/GM/NIGMS NIH HHS/United States
PT  - Journal Article
DEP - 20190728
PL  - Switzerland
TA  - Data (Basel)
JT  - Data
JID - 101699766
PMC - PMC7377353
MID - NIHMS1608031
OTO - NOTNLM
OT  - FcepsilonRI
OT  - IgE receptor
OT  - RBL-2H3
OT  - TIRF
OT  - plasma membrane
OT  - rat basophilic leukemia cells
OT  - supported lipid bilayer
OT  - total internal reflection fluorescence microscopy
COIS- Conflicts of Interest: The authors declare no conflict of interest. The funders 
      had no role in the design of the study; in the collection, analyses, or 
      interpretation of data; in the writing of the manuscript, or in the decision to 
      publish the results.
EDAT- 2020/07/25 06:00
MHDA- 2020/07/25 06:01
PMCR- 2020/07/23
CRDT- 2020/07/25 06:00
PHST- 2020/07/25 06:00 [entrez]
PHST- 2020/07/25 06:00 [pubmed]
PHST- 2020/07/25 06:01 [medline]
PHST- 2020/07/23 00:00 [pmc-release]
AID - 111 [pii]
AID - 10.3390/data4030111 [doi]
PST - ppublish
SO  - Data (Basel). 2019 Sep;4(3):111. doi: 10.3390/data4030111. Epub 2019 Jul 28.