Morphological and Physiological Monitoring of Tumor Spheroids Using Optical Coherence Tomography

Samenvatting

Source: Huang, Y. et. al., Longitudinal Morphological and Physiological Monitoring of Three-dimensional Tumor Spheroids Using Optical Coherence Tomography. J. Vis. Exp. (2019)

This video demonstrates the use of optical coherence tomography (OCT) to monitor tumor spheroids in a multi-well plate by capturing high-resolution cross-sectional images. By analyzing light reflections from cellular structures, OCT constructs 3D models to assess morphological features like size and shape while also distinguishing viable from non-viable cells based on light scattering changes over time.

Protocol

1. Preparation of Cells

Resultaten

Materialen

Lijst van materialen gebruikt in dit artikel

Naam	Bedrijf	Catalogusnummer	Opmerkingen
Custom Spectral Domain OCT imaging system			Developed in our lab
Superluminescent diode (SLD)	Thorlabs	SLD1325	Light source
2×2 single mode fused fiber coupler, 50:50 splitting ratio	AC Photonics	WP13500202B201
Reference Arm
Lens Tube	Thorlabs
Adapter	Thorlabs
Collimating Lens	Thorlabs	AC080-020-C
Focusing Lens	Thorlabs
Kinematic Mirror Mount	Thorlabs
Mirror	Thorlabs
1D Translational Stage	Thorlabs
Continuous neutral density filter	Thorlabs
Pedestrial post	Thorlabs
Clamping fork	Thorlabs
Sample arm
Lens tube	Thorlabs
Adapter	Thorlabs
Collimating lens	Thorlabs	AC080-020-C
Galvanometer	Thorlabs
Relay lens	Thorlabs	AC254-100-C	Two Relay lens to make a telescope setup
Triangle mirror mount	Thorlabs
Mirror	Thorlabs
Objective	Mitutoyo
Pedestrial post	Thorlabs
Clamping fork	Thorlabs
Polarization controller	Thorlabs
30mm cage mount	Thorlabs
Cage rod	Thorlabs
Stage
3D motorized translation stage	Beijing Mao Feng Optoelectronics Technology Co., Ltd.	JTH360XY
2D Tilting stage
Rotation stage
Plate holder			3D printed
Spectrometer
Lens tube	Thorlabs
Adapter	Thorlabs
Collimating lens	Thorlabs	AC080-020-C
Grating	Wasatch		G = 1145 lpmm
F-theta lens	Thorlabs	FTH-1064-100
InGaAs line-scan camera	Sensor Unlimited	SU1024-LDH2
Cell Culture Component
HCT 116 Cell line	ATCC	CCL-247
Cell culture flask	SPL Life Sciences	70025
Pipette	Fisherbrand	14388100
Pipette tips	Sorenson Bioscience	10340
Gibco GlutaMax DMEM	Thermo Fisher Scientific	10569044
Fetal bovine serum, certified, US origin	Thermo Fisher Scientific	16000044
Antibiotic-Antimycotic (100X)	Thermo Fisher Scientific	15240062
Corning 96-well clear round bottom ultra-low attachment microplate	Corning	7007
Gibco PBS, pH 7.4	Thermo Fisher Scientific	10010023
Gibco Trypsin-EDTA (0.5%)	Thermo Fisher Scientific	15400054
Forma Series II 3110 Water-Jacketed CO2 incubator	Thermo Fisher Scientific	3120
Gloves	VWR	89428-750
Parafilm	Sigma-Aldrich	P7793
Transfer pipets	Globe Scientific	138080
Centrifuge	Eppendorf	5702 R	To centrifuge the 15 mL tube
Centrifuge	NUAIRE	AWEL CF 48-R	To centrifuge the 96-well plate
Microscope	Olympus
List of Commerical OCTs
SD-OCT system	Thorlabs	Telesto Series
SD-OCT system	Wasatch Photonics	WP OCT 1300 nm
Software for Data Analyses
Basic Image Analysis	NIH	ImageJ	Fiji also works.
3D Rendering	Thermo Fisher Scientific	Amira	Commercial software. Option 1
3D Rendering	Bitplane	Imaris	Commercial software. Option 2. Used in the protocol
OCT acquisition software			custom developed in C++.
Stage control	Beijing Mao Feng Optoelectronics Technology Co., Ltd.	MRC_3	Incorporated into the custom OCT acquisition code
OCT processing software			Custom developed in C++. Utilize GPU. Incorporated into the custom OCT acquisition code.
Morphological and Physiological Analysis			Custom developed in MATLAB