An optimum study on the laser scanning confocal microscopy techniques for BiFC assay using plant protoplast

Botanical Studies

Table 1 Case studies of protein interactions using the protoplast BiFC system

Color	Fluorescence protein	Excitation (nm)	Emission (nm)	Microscopy	Protoplast source	Observation of chloroplast autofluorescence	Nucleus localization	References
Red	RFP	549	570	LSCM	Tobacco BY2	No	–	Jach et al. (2006)
Red	RFP	558	583	LSCM	Tobacco	No	BF/GFP	Li et al. (2012)
Yellow	YFP	514	530–560	LSCM	Rice	Yes	BF	Zhang et al. (2011)
	YFP	480–520	505–565	FL	Parsley	No	CFP	Stolpe et al. (2005)
	YFP	488	505–530	FL	Arabidopsis	Yes	BF	Wu et al. (2009)
	YFP	Single laser	–	LSCM	Tobacco	Yes	BF	Citovsky et al. (2006)
	EYFP	480	500–550	LSCM	Arabidopsis root	Yes	mCherry/BF	Dervisi et al. (2022)
	EYFP	488	505–565	LSCM	Arabidopsis	No	BF	Olejnik et al. (2011)
	YFP	514	–	LSCM	Arabidopsis	Yes	BF	Zhang et al. (2016)
	YFP	480	500–550	LSCM	Arabidopsis	No	BF	Hussain et al. (2018)
	YFP	–	–	LSCM	Arabidopsis	Yes	DAPI	Chang et al. (2019)
	nVenus + cCFP	–	–	LSCM	Arabidopsis	Yes	DAPI/BF	Liu et al. (2021)
	nVenus + cCFP	488	500–530	FL/LSCM	Tobacco BY2	No	mCherry/BF	Lee et al. (2008)
Green	GFP	450–490	520–560	FL	Rice	Yes	BF	Chen et al. (2006)
	GFP	–	450–490	FL/LSCM	Freesia/Arabidopsis	No	RFP	Shan et al. (2019)
	GFP	488	500–530	LSCM	Rice	Yes	BF	Zhang et al. (2011)
	GFP	–	–	LSCM	Arabidopsis	Yes	DAPI/BF	Liu et al. (2021)
Cyan	nCerulean + cCFP	458	480–520	FL/LSCM	Tobacco BY2	No	mCherry/BF	Lee et al. (2008)
Cyan	nCerulean + cCFP	–	–	LSCM	Arabidopsis root	Yes	BF	Dervisi et al. (2022)

LSCM laser scanning confocal microscope, FL fluorescence microscope, BF bright field