Table 2 Experimental results reveal an optimization tradeoff between trackable diffusion rates and localization error for MFX-enabled single particle tracking

\({{\rm{S}}}\)	PL	DMP	\({{{\rm{t}}}}_{{{\rm{dwell}}}}\)	\({{{\rm{D}}}}_{{{\rm{eMSD}}}}\) \([{{\rm{\mu }}}{{{\rm{m}}}}^{2}/{{\rm{s}}}]\)	\({{{\rm{\sigma }}}}_{{{\rm{eMSD}}}}\) \(\left[{{\rm{nm}}}\right]\)	\({{{\rm{\sigma }}}}_{{{\rm{CRLB}}}}\) \([{{\rm{nm}}}]\)	\(\left\langle {{\rm{\eta }}}\right\rangle\)	\(\left\langle {{{\rm{t}}}}_{{{\rm{loc}}}}\right\rangle\) \([{{\rm{\mu }}}{{\rm{s}}}]\)	\(\frac{\left\langle {{\rm{N}}}\right\rangle }{\left\langle {{\rm{\eta }}}\right\rangle {{{\rm{N}}}}_{{{\rm{PL}}}}}\)	\({{\rm{n}}}\)
I	10	0	100	2.46	30.30	20.03	1	150	2.13	855
II	10	5	100	0.07	16.52	20.03	1	150	4.10	924
II	10	2	100	0.49	15.53	20.03	1	150	4.49	988
II	10	0	100	0.63	28.02	20.03	1	150	3.36	1075
II	10	0	100	0.63	27.96	20.03	1	150	3.36	1061
II	20	0	100	0.52	22.78	14.35	1	151	2.27	841
II	50	0	100	0.48	14.05	9.08	2	282	0.71	534
III	10	0	100	0.46	30.37	20.03	1	150	1.53	576
III	20	0	100	0.36	23.51	14.35	2	281	0.67	325
III	50	0	100	0.23	13.46	9.08	5	676	0.22	323
II	50	0	50	0.28	4.67	9.08	5	437	0.23	779
II	50	0	100	0.28	5.50	9.08	3	414	0.42	763
II	50	0	200	0.21	10.91	9.08	2	483	0.82	564
II	TIRF SPT			0.64	39.86	-	1	14925	-	1810

In addition to the ideal CRLB \({\sigma }_{{CRLB}}\left(\bar{r}=\bar{0}\right)\) (see Supplementary Material of ref. ⁸, p. 11) we list the lateral diffusion coefficient \({D}_{{eMSD}}\) as well as the lateral localization error \({\sigma }_{{eMSD}}\) extracted from the ensemble-average Mean-Squared-Displacement (eMSD) using Optimal Least Squares Fitting (OLSF) assuming momentary Brownian motion. Here \(\eta\) refers to the number of cycles, \({t}_{{loc}}\) is the time-to-localization, \(N\) is the number of photons, \({N}_{{PL}}\) is the PL, and \(\left\langle \ldots \right\rangle\) the scale-appropriate average value. We indicate the sample size \(n\), i.e., the number of tracks per set, in the last column. The first column \(S\) indicates the sample in reference: Dye on GUV-patch SLB (I), QDs on GUV-patch SLB (II), QDs on Lipid-Deposition-SLB (III).

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