MALC, a novel microinjection method for loading of macromolecules into cultured neurons.
Microinjection is one of the most useful and important methods in cellular neurobiology. However, direct insertion and retraction of a capillary produce physical stresses inside cells that make it difficult to apply on small and fragile cells, especially the central nervous system (CNS) neurons. In this study, we developed a novel method called MALC (microinjection method assisted with laser activated chromophore) to avoid this disadvantage of the conventional method. In MALC, a capillary is just attached on cell surface, and combination of a newly developed photosensitizer with a laser pointing system makes the plasma membrane permeable to molecules. This novel method can expand the application of the microinjection method and realize molecular manipulations inside CNS neurons.
Yano
Ryoji
R
Laboratory for Cellular Information Processing, The Brain Science Institute (BSI), RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.
Okada
Daisuke
D
Yap
Chan Choo
CC
Mabuchi
Kunihiko
K
Muguruma
Hitoshi
H
Saito
Takashi K
TK
D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D002462:Cell Membrane / Q000502:physiology; D002478:Cells, Cultured; D004247:DNA / Q000494:pharmacology; D005456:Fluorescent Dyes / Q000494:pharmacology*; D006624:Hippocampus / Q000166:cytology; D007834:Lasers; D008564:Membrane Potentials / Q000502:physiology; D008845:Microinjections / Q000295:instrumentation / Q000379:methods*; D009474:Neurons / Q000166:cytology / Q000502:physiology*; D009930:Organic Chemicals; D010777:Photochemistry; D012333:RNA, Messenger / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D013314:Stress, Mechanical
D000906:Antibodies; D005456:Fluorescent Dyes; D009930:Organic Chemicals; D012333:RNA, Messenger; C071336:calcium green; D004247:DNA; D002118:Calcium