Real-time hyperspectral fluorescence imaging of pancreatic β-cell dynamics with the image mapping spectrometer (IMS)

Abstract

SummaryThe development of multi-colored fluorescent proteins, nanocrystals and organic fluorophores, along with the resulting engineered biosensors, has revolutionized the study of protein localization and dynamics in living cells. Hyperspectral imaging has proven to be a useful approach for such studies, but this technique is often limited by low signal and insufficient temporal resolution. Here, we present an implementation of a snapshot hyperspectral imaging device, the image mapping spectrometer (IMS), which acquires full spectral information simultaneously from each pixel in the field without scanning. The IMS is capable of real-time signal capture from multiple fluorophores with high collection efficiency (,65%) and image acquisition rate (up to 7.2 fps).

Snapshot 3D optical coherence tomography system using image mapping spectrometry

“…The earlier mappers used in IMS were fabricated using a raster-fly cutting technique on a four-axis Nanotech Ultra Precision milling machine [24]. Here we used a ruling technique which has been shown recently to exhibit several advantages over raster-fly cutting [30]. The raster-fly cutting process is significantly slower than ruling, and it creates a large inconsistency in facet widths.…”

A review of snapshot multidimensional optical imaging: Measuring photon tags in parallel

“…With a large-format CCD, a current state-of-the-art IMS can measure a 350 × 350 × 48 ( x,y ,λ) datacube [45] within a single camera snapshot. The IMS has been demonstrated in combination with a variety of imaging modalities, such as microscopy [16, 24, 46–48], endoscopy [45], fundus photography [49], and macroscopy [50, 51], and has been employed for imaging both in the visible [45, 46] and infra-red spectral ranges [51]. …”

Lenslet array tunable snapshot imaging spectrometer (LATIS) for hyperspectral fluorescence microscopy

“…These spectral imagers can be designed to be advantageous over existing scanning spectrometers due to longer pixel dwell times, permitting their use in dim applications, and higher speeds, which eliminate motion artifacts for dynamic scenes [18]. The advantages of these spectral imagers have seen their increased use in microscopy applications over the last few years [19,20] and in vivo tissue imaging [21][22][23][24][25]. Each pixel's exposure time to the collected light is the same as the time used to acquire the datacube, which increases irradiance per pixel in comparison to scanning techniques with the same datacube acquisition time or frame rate.…”