TY - JOUR
T1 - Characteristics of genetic tags for correlative light and electron microscopy
AU - Beatty, Kimberly E.
AU - López, Claudia S.
N1 - Publisher Copyright:
© 2023 Elsevier Ltd
PY - 2023/10
Y1 - 2023/10
N2 - Fluorescence microscopy is indispensable in live cell studies of fluorescently-labeled proteins, but has limited resolution and context. Electron microscopy offers high-resolution imaging of cellular ultrastructure, including membranes, organelles, and other nanoscale features. However, identifying proteins by EM remains a substantial challenge. There is potential to combine the strengths of both FM and EM through correlative light and EM (CLEM), and bridging the two modalities enables new discoveries and biological insights. CLEM enables cellular proteins to be observed dynamically, across size scales, and in relationship to sub-cellular structures. A central limitation to using CLEM is the scarcity of methods for labeling proteins with CLEM reporters. This review will describe the characteristics of genetic tags for CLEM that are available today, including fixation-resistant fluorescent proteins, 3,3′-diaminobenzidine (DAB)-based tags, metal-chelating tags, DNA origami tags, and VIP tags.
AB - Fluorescence microscopy is indispensable in live cell studies of fluorescently-labeled proteins, but has limited resolution and context. Electron microscopy offers high-resolution imaging of cellular ultrastructure, including membranes, organelles, and other nanoscale features. However, identifying proteins by EM remains a substantial challenge. There is potential to combine the strengths of both FM and EM through correlative light and EM (CLEM), and bridging the two modalities enables new discoveries and biological insights. CLEM enables cellular proteins to be observed dynamically, across size scales, and in relationship to sub-cellular structures. A central limitation to using CLEM is the scarcity of methods for labeling proteins with CLEM reporters. This review will describe the characteristics of genetic tags for CLEM that are available today, including fixation-resistant fluorescent proteins, 3,3′-diaminobenzidine (DAB)-based tags, metal-chelating tags, DNA origami tags, and VIP tags.
KW - Correlative light and electron microscopy
KW - Electron microscopy
KW - Fluorescence microscopy
KW - Fluorescent protein
KW - Genetic tag
KW - Imaging
UR - http://www.scopus.com/inward/record.url?scp=85164746505&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85164746505&partnerID=8YFLogxK
U2 - 10.1016/j.cbpa.2023.102369
DO - 10.1016/j.cbpa.2023.102369
M3 - Review article
C2 - 37453163
AN - SCOPUS:85164746505
SN - 1367-5931
VL - 76
JO - Current Opinion in Chemical Biology
JF - Current Opinion in Chemical Biology
M1 - 102369
ER -