Fisher information and the Cramér–Rao lower bound in single-pixel localization microscopy with spatiotemporally modulated illumination

Single-pixel imaging, the concept that an image can be captured via a single-pixel detector, is a cost-effective yet powerful technique to reduce data acquisition duration without sacrificing image resolution when properly structured illumination patterns are introduced. Normally, the image reconstruction process is subject to the diffraction limit. Here, we study the possibility of exploiting the information contained in the illumination patterns to enable a form of single-pixel localization microscopy (SPLM) for super-resolution. This concept is inspired by coherent holographic image reconstruction by phase transfer (CHIRPT) microscopy. CHIRPT microscopy is a single-pixel imaging technique that uses structured illumination that is spatiotemporally modulated (STM) so that a unique temporal modulation pattern is imparted to each point within a large illumination volume. The fluorescent light emitted by molecules contains the same temporal modulations as the illumination patterns at the locations of the molecules. By recording a portion of the total emitted fluorescent power, the signal may be numerically processed to form an image. Unique temporal modulation patterns that excite fluorescent probes at each point can also be used to localize individual molecules by matching their particular temporal light emission patterns to the measured temporal signal. This paper evaluates the feasibility of SPLM with STM illuminations used in and inspired by CHIRPT microscopy via the information content its data carry about the emitter location(s). More specifically, we provide the mathematical formalism of Fisher information (FI) and the Cramér–Rao lower bound (CRLB) associated with the location parameters of the emitter(s). The FI and CRLB are then numerically evaluated under different experimental assumptions to assess the effects of experimental parameters on localization precision. Last, we compare the single-pixel CRLB to that from camera-based single-molecule localization microscopy in the localization of a single emitter. We show that SPLM has several distinguishing characteristics that provide certain advantages, such as relatively constant CRLB over a very large illumination volume and improved CRLB for 3D localization due to the information coupling introduced by simultaneous modulations of the transverse axes.