![Sample our Bioscience journals, sign in here to start your access, Latest two full volumes FREE to you for 14 days]({"type":"image" , "src" : "/sda/5925/TOC-Subject-Sample-2021-Bioscience.jpg"})
ABSTRACT
Each cell produces its own responses even if it appears identical to other cells. To analyze these individual cell characteristics, we need to measure trace amounts of molecules in a single cell. Nucleic acids in a single cell can be easily amplified by polymerase chain reaction, but single-cell measurement of proteins and sugars will require de novo techniques. In the present study, we outline the techniques we have developed toward this end. For proteins, our ultrasensitive enzyme-linked immunosorbent assay (ELISA) coupled with thionicotinamide-adenine dinucleotide cycling can detect proteins at subattomoles per assay. For sugars, fluorescence correlation spectroscopy coupled with glucose oxidase-catalyzed reaction allows us to measure glucose at tens of nM. Our methods thus offer versatile techniques for single-cell-level analyses, and they are hoped to strongly promote single-cell biology as well as to develop noninvasive tests in clinical medicine.
Abbreviations
| 3α-HSD | = | 3α-hydroxysteroid dehydrogenase |
| AIDS | = | acquired immune deficiency syndrome |
| ALP | = | alkaline phosphatase |
| BSA | = | bovine serum albumin |
| ELISA | = | enzyme-linked immunosorbent assay |
| FCS | = | fluorescence correlation spectroscopy |
| GOD | = | glucose oxidase |
| HRP | = | horseradish peroxidase |
| PCR | = | polymerase chain reaction |
| thio-NAD | = | thionicotinamide-adenine dinucleotide |
| tyramide-TMR | = | tyramide labeled with tetramethyl rhodamine |
Disclosure of potential conflicts of interest
SW is an employee of BL Co., Ltd., and MM and KN are employees of TAUNS Laboratories, Inc.. MK, AN, MN, TY, TM, and EI declare that they have no conflicts of interest.
Funding
This study was supported by a grant for the Development of Systems and Technology for Advanced Measurement and Analysis from JST, by a grant for the Regional Innovation Strategy Support Program 2009 from MEXT, by a grant for the New Regional Consortium Research and Development Project from METI, and by the Hokkaido Bureau of Economy, Trade, and Industry to E.I. and the others.