Sara Abrahamsson: "Latest developments in multifocus microscopy: deeper and faster high resoluti..."

Описание: Sara Abrahmsson (UCSC) presents "Latest developments in multifocus microscopy: deeper and faster high resolution simultaneous 3D imaging."

Abstract:
My research group at UCSC is happy to introduce to you our new imaging system M25 (in preparation for publication). This new microscope allows truly simultaneous high-resolution high-speed imaging in a 3D volume consisting of 25 focal planes, each recorded on a fast (up to 160Hz), tiny and sensitive camera in a 5x5 camera array. M25 is based on aberration-corrected multifocus microscopy (MFM), which I developed during my Ph.D. for Dr Mats Gustafsson at UCSF and built and tested at HHMI Janelia (Abrahamsson et al. Nat Methods 2012). MFM technology is based on the use of specially designed diffractive Fourier optics, which have unique capabilities that enable multiplexing and refocusing of the light from a wide-field microscope, to enable truly simultaneous imaging of entire 3D volumes at diffraction limited resolution. MFM imaging is also compatible with localization and super-resolution microscopy. The aim of the M25 instrument is to allow biologists to image fast dynamic processes, such as neuronal signaling, in entire small living model organisms in 3D, at high speed and with high resolution. M25 works both in fluorescent and bright-field mode, thus also allowing label free imaging, which can be a minimally invasive imaging method.

0:00 Introduction
1:58 Optical Systems Invention and Design for 3D Microscopy
3:06 Microscopy is an unfortunate series of compromises
4:03 Diffractive Optics Nanofabrication
5:40 Bespoke Microscope Commissioning
6:46 The 2D Image
8:04 Light-Field Microscopy: "multiple stereo" imaging - loses resolution
8:50 Diffractive Fourier optics: Harnessing the wave nature of light
9:23 Step 1: Multiplex the image
10:32 Optimizing the grating function
11:00 Pixelflipper algorithm
12:38 Step 2: Distort the grating
16:33 MFM transcription imaging at Janelia, a long time ago
17:11 Functional Neuronal Imaging Bargmann Lab, Rockefeller University
17:55 Design a grating function for 25 planes
18:19 M25 in monochromatic brightfield microscopy
21:33 Built by my very first PhD student Dr Eduardo Hirata
22:57 Proof of Concept M25: Snap shots of model organisms Lamprey and Zebrafish
23:41 C. elegans muscle activity
25:03 SIM improves both 3D resolution and contrast
25:10 Spatial frequency mixing
25:25 SIM data reconstruction
27:43 My 5 cents: We need commercial microscopes
31:50 M25 PSF and some dancing bead tracking
38:02 Radical new design in camera array prism-less dispersion correction: "M25"
40:36 89% photon efficient multiplexing with a multi-layer phase grating