Reduction in nonfluorescence state of quantum dots on an immunofluorescence staining.
Fluorescence quantum dots are widely used in immunofluorescence staining because of their intense and stable fluorescence. However, the nonfluorescence state of the quantum dots is their disadvantage. Here, the nonfluorescence state of the dots labeled to cells and tissues was suppressed. Cells and tissues where the receptor HER2 had been overexpressed were fixed and then labeled with anti-HER2 crosslinked with the dots. The intensity of the dots increased with the illumination time. The majority of the single dots were in the nonfluorescence state at beginning of the illumination period and the number of fluorescence dots observed increased with the illumination time. Living cells were also labeled with the anti-HER2-Qdots. Blinking and bleaching of the Qdots was effectively suppressed by adding beta-mercaptoethanol and glutathione. Therefore, the movement of the Qdots bound to cell membrane could be observed for long periods of time.
Li-Shishido
Songhua
S
Division of Surgical Oncology, Graduate School of Medicine, Tohoku University, Japan.
Watanabe
Tomonobu M
TM
Tada
Hiroshi
H
Higuchi
Hideo
H
Ohuchi
Noriaki
N
D000818:Animals; D001943:Breast Neoplasms / Q000378:metabolism* / Q000473:pathology*; D002460:Cell Line; D005260:Female; D005455:Fluorescent Antibody Technique / Q000379:methods*; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D008856:Microscopy, Fluorescence / Q000379:methods*; D009363:Neoplasm Proteins / Q000378:metabolism*; D045663:Quantum Dots; D013194:Staining and Labeling / Q000379:methods
D009363:Neoplasm Proteins