High sensitivity detection of protein molecules picked up on a probe of atomic force microscope based on the fluorescence detection by a total internal reflection fluorescence microscope.
We developed a method to detect and identify proteins on a probe of the atomic force microscope (AFM) with a high sensitivity. Due to a low background noise of the total internal reflection fluorescence microscope employed as a detecting system, we were able to achieve a high enough sensitivity to detect zeptomole orders of protein molecules immobilized on the tip. Several different methods to immobilize protein molecules to AFM-probes were tested, meant for a wide range of applications of this method. Furthermore, we demonstrated that different proteins were clearly distinguished by immunofluorescence microscopy on the probe using their specific antibodies.
Yamada
Takafumi
T
Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 Nagatsuta-cho, Midoriku, Yokohama 226-8501, Japan.
Afrin
Rehana
R
Arakawa
Hideo
H
Ikai
Atsushi
A
D000906:Antibodies; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000737:chemistry* / Q000235:genetics / Q000648:ultrastructure; D018625:Microscopy, Atomic Force / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods; D016297:Mutagenesis, Site-Directed; D015202:Protein Engineering; D011506:Proteins / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry / Q000648:ultrastructure; D012680:Sensitivity and Specificity
D000906:Antibodies; D008164:Luminescent Proteins; D011506:Proteins; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins