Reagent or Resource	Source	Identifier
Antibodies
Rat Anti-mTFP	Cai Lab
Chicken Anti-GFP	Cai Lab
Rabbit Anti-mNeonGreen	Cai Lab
Mouse Anti-mKusabira-Orange2	Cai Lab
Guinea Pig Anti-mKate2	Cai Lab
Alexa Fluor 594 Donkey Anti-Rat	Life Technologies	A21209
Alexa Fluor 488 Donkey Anti-Chicken	Jackson ImmunoResearch	703-545-155
Atto 490LS Goat Anti-Rabbit	Hypermol	2309
CF 555 Donkey Anti-Mouse	Sigma	SAB4600060
CF 633 Donkey Anti-Guinea Pig	Sigma	SAB4600129
Alexa Fluor 647 Donkey Anti-Guinea Pig	Jackson ImmunoResearch	706-605-148
Bacterial and Virus Strains
Mach1-T1 Chemically Competent E. coli	Thermo Scientific	C862003
Stbl3 Chemically Competent E. coli	Thermo Scientific	C737303
Chemicals, Peptides, and Recombinant Proteins
Acrylic acid N-hydroxysuccinimide ester	Sigma	A8060
Sodium Acrylate	Sigma	408220
Acrylamide	Sigma	A9099
N,N’-methylenebisacrylamide	Sigma	M7279
Tetramethylethylenediamine (TEMED)	Sigma	T7024
Ammonium Persulfate	Sigma	A3678
4-hydroxy-2,2,6, 6-tetramethylpiperidin-1-oxyl (4HT)	Sigma	176141
Startingblock (PBS) blocking buffer	Thermo Scientific	37578
Proteinase K	NEB	P8107
Experimental Models: Cell Lines
D. melanogaster: S2	Bing Ye Lab
M. musculus: Neuro-2a	ATCC	CCL-131
Experimental Models: Organisms/Strains
D. melanogaster: w1118;;	BDSC	RRID: BDSC_5905
D. melanogaster: hsFlp112;;	BDSC
D. melanogaster: hsFlp112;Sp/CyO;TM2,Ubx/TM6B,Tb	Bing Ye Lab
D. melanogaster: hsFlp112;;elav-Gal4	This study
D. melanogaster: hsFlp112;TRH-Gal4;	This study
D. melanogaster: yw;;mBitbow1.0	This study
D. melanogaster: yw;;nBitbow1.0	This study
D. melanogaster: yw;gBitbow1.0;	This study
D. melanogaster: yw;gBitbow1.0/CyO;mBitbow1.0, nBitbow1.0	This study
D. melanogaster: yw;;mBitbow2.0	This study
D. melanogaster: yw;;mBitbow2.1	This study
D. melanogaster: yw;;2x[mBitbow2.0]	This study
D. melanogaster: yw;;2x[mBitbow2.1]	This study
D. melanogaster: w;elav-Gal4/CyO;	BDSC	8765
D. melanogaster: w;TRH-Gal4;	BDSC	38388
D. melanogaster: w;;R53C10-Gal4	BDSC	38873
D. melanogaster: w;;R67A06-Gal4	BDSC	39397
Recombinant DNA
pmAmetrine-N1	Addgene	54505
pmTFP1-N1	Addgene	54521
pmNeonGreen-N1	Allele Biotechnology
pmKusabira-Orange2-N1	Sakaue-Sawano et al., 2008
ptdKatushka2-N	Shcherbo et al., 2009
pmCitrine-N1	Addgene	54594
pmCherry-N1	Shaner et al., 2004
pJFRC-MUH	Addgene	26213
pattB-synaptobrevin-GAL4-hsp70	Addgene	46107
pJFRC81-10XUAS-IVS-Syn21-GFP-p10	Addgene	36432
pMT-Gal4	DGRC	1042
pUAST-Flp	DGRC	1020
pCAG-FlpINT	Dawen Cai Lab
pDC-MUH	This study
pDC-MUH-p10pA	This study
pCMV-2xmAmetrine-N	This study
pDC-UAS-mBitbow1.0	This study
pDC-UAS-nBitbow1.0	This study
pDC-UAS-gBitbow1.0	This study
pDC-UAS-mBitbow2.0	This study
pDC-UAS-mBitbow2.1	This study
pAc5-Flp-p10pA	This study
pCMV-3FRT-mCherry-F14-mTFP	This study
pCMV-3FRT-mCherry-F15-mCit	This study
pCMV-3FRT-F14-mTFP-F15-mCit	This study
pCMV-3FRT-F14-F15-mCit-F13-mCherry	This study
pCAG-Flpbow3	Cai et al., 2013
Software and Algorithms
Fiji	Schindelin et al., 2012	https://fiji.sc/
nTracer (Fiji plugin)	Roossien et al., 2019	https://www.cai-lab.org/ntracer-tutorial
Spectral Unmixing (Fiji plugin)	Joachim Walter	https://imagej.nih.gov/ij/plugins/spectral-unmixing.html
Lasergene	DNASTAR	https://www.dnastar.com/
Graphpad Prism	Graphpad Software	https://www.graphpad.com/
Vaa3D	Peng et al., 2010	http://home.penglab.com/proj/vaa3d/home/
Blender	Blender Foundation	https://www.blender.org/
NumPy	NumPy.org	https://numpy.org/
umap-learn	McInnes et al., 2018	https://github.com/lmcinnes/umap