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Nano Convergence

Table 1 Summary of ExM methods based on the target biomolecules

From: Nanoscale fluorescence imaging of biological ultrastructure via molecular anchoring and physical expansion

Target biomolecule

Method name

Chemical anchor(s)

Crosslinker(s)

Expansion factor (single round unless otherwise noted)

Commercial availability of the anchor(s)

References

Proteins

proExM

Acryloyl-X, SE

N,N′-Methylenebisacrylamide

~ 4–4.5×

Yes

[37]

ExM compatible with conventional fluorescently labeled antibodies and fluorescent proteins

Methacrylic acid N-hydroxysuccinimide ester (MA-NHS), glutaldehyde (GA)

N,N′-Methylenebisacrylamide

~ 4×

Yes

[38]

MAP

Acrylamide and formaldehyde

N,N′-Methylenebisacrylamide

~ 4×

Yes

[45]

Original ExM/iExM

Acrydite-modified oligonucleotides

N,N′-Methylenebisacrylamide, N,N′-(1,2-dihydroxyethylene) bisacrylamide (DHEBA), N,N′-bis(acryloyl)cystamine (BAC)

~ 4.5–5.5×, ~ 16–22× (two rounds), ~ 53× (three rounds)

No

[34, 46]

pan-ExM

Acrylamide and formaldehyde

N,N′-(1,2-dihydroxyethylene) bisacrylamide (DHEBA), N,N′-methylenebisacrylamide

~ 3.8–4×, ~ 13–21× (two rounds)

Yes

[47]

TREx

Acryloyl-X, SE

N,N′-Methylenebisacrylamide

~ 10×

Yes

[48]

X10 ExM

Acryloyl-X, SE

N,N-Dimethylacrylamide (DMAA)

~ 9.5×–11.5×

Yes

[49, 50]

NIFS hydrogel

MA-NHS

N,N-Ethylenebisacrylamide (EBIS)

~ 9×

Yes

[51]

ExR

Acryloyl-X, SE

N,N′-methylenebisacrylamide

~ 4×, ~ 20× (two rounds)

Yes

[52]

Tetra-gel (TG)

NHS-azide

Step-growth polymerization (no crosslinking); the polymerization proceeds via click reaction between the azide and alkyne groups of the monomers

~ 3.0–3.5×, ~ 10–16× (two rounds), ~ 40× (three rounds)

Yes

[53, 54]

LR-ExM

Trifunctional anchors (HOOC-MA-biotin, HOOC-MA-DIG, BG-MA-biotin, BG-MA-DIG, and BC-MA-DIG)

N,N′-Methylenebisacrylamide

~ 4.3–4.7×

No

[55]

PhASE-ExM

Acryloyl-X, SE

8-arm and 10 kDa PEG-SH, N,N′-methylenebisacrylamide

~ 3.25×

Yes

[56]

U-ExM

Acrylamide and formaldehyde

N,N′-Methylenebisacrylamide

~ 4×

Yes

[57, 58]

Nucleic acids

ExFISH

Label X

N,N′-Methylenebisacrylamide

~ 3.3×

No (Synthesized from Label-IT and AcX)

[59]

EASI-FISH

MelphaX

N,N′-Methylenebisacrylamide

~ 2–3×

No (Synthesized from Melphalan and AcX)

[60]

ExSeq

Label X

N,N′-Methylenebisacrylamide

~ 4×

No (Synthesized from Label-IT and AcX)

[61]

Lipids

mExM

Membrane probe pGk5b

N,N′-Methylenebisacrylamide

~ 3×

No

[62]

LExM

Trifunctional LExM reagent

N,N′-Methylenebisacrylamide

~ 4–8×

No

[63]

Sphingolipid ExM

α–NH2–ω–N3–C6–ceramide and GA

N,N′-Methylenebisacrylamide

 ~ 4 and 10×

No

[64]

Multiplexed labeling

TRITON

Trivalent TRITON linker

N,N′-Methylenebisacrylamide

~ 3.2×

No

[40, 65]

Click-ExM

Interaction between biotin, streptavidin, and Acryloyl-X, SE (or GA)

N,N′-Methylenebisacrylamide

~ 4.5×

No

[66]

Physical entanglement

eMAP

Entanglement by the polymer chains (no covalent anchoring)

N,N′-Methylenebisacrylamide

~ 4 and 10×

Yes

[67]

ELAST

Entanglement by the polymer chains (no covalent anchoring)

N,N′-Methylenebisacrylamide

~ 2–3×

Yes

[68]

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