A Do-It-Yourself and 3D-Printed Time-Lapse Imaging Platform for In-Incubator Live Cell Observation

Summary

This protocol describes a do-it-yourself (DIY), low-cost, three-dimensional (3D)-printed time-lapse imaging platform controlled by Raspberry Pi, built for use inside standard incubators. It uses programmable light-emitting diode (LED) lighting to achieve bright field or oblique imaging and runs open-source Python software for automated time-lapse workflows.

Abstract

Integrated in-incubator microscopy systems and all-in-one time-lapse culture incubators are often expensive and proprietary, which limits accessibility and customization. Open-source alternatives exist but often involve tradeoffs in imaging performance, illumination control, or adaptability. Here, we describe a do-it-yourself (DIY), low-cost, three-dimensional (3D)-printed time-lapse imaging platform designed for use inside standard cell culture incubators. The device employs a Raspberry Pi–based controller with stepper motor actuation and a programmable light-emitting diode (LED) backlight that enables both bright-field and oblique illumination modes. The system supports automated multi-well imaging, scheduled time-lapse acquisition, and real-time preview through open-source Python software. Performance validation demonstrated reliable long-term imaging of cultured cells and early embryos, with stable operation under incubator conditions. Optical resolution testing using a United States Air Force (USAF) 1951 target confirmed a minimum resolvable feature of approximately 1.55 µm. Motion evaluation showed low drift and high repeatability, with positioning accuracy closely matching commanded displacements. The system is compatible with multiple plate formats and can be adapted for specialized culture setups. Overall, this platform provides a cost-effective, reproducible, and customizable solution for long-term live-cell and embryo imaging in laboratory settings.

Introduction

Live cell imaging is widely applied across biological research to investigate dynamic cellular processes, including cell proliferation^1,2, differentiation³, migration⁴, and, notably, early embryonic development for embryo quality assessment in IVF laboratories^5,6. Existing solutions primarily include chamber-based microscope systems⁷ and integrated time-lapse incubators⁸. Although these platforms enable extended time-lapse imaging under controlled environmen....

Protocol

The tumor cell lines and animal embryos were obtained from the NHC Key Laboratory of Chronobiology (Sichuan University) and were handled in accordance with the protocols approved by the local institutional review board and ethics committee, as well as relevant national regulations. This method must not be applied directly in clinical practice or used for human embryo culture without sufficient experimental validation.

NOTE: The hardware (PCB design files and 3D models/STL files) is released under the CERN Open Hardware License Version 2 – Strongly Reciprocal (CERN-OHL-S-2.0), and the software (Controller desktop GUI and Raspberry Pi backend....

Results

Resolution testing
To evaluate the optical resolution of the imaging module, a USAF 1951 resolution target was imaged under bright-field illumination using a 10× objective. Raw images were acquired at 3840 × 2160 pixels. As shown in Figure 6A, the line-pair patterns were clearly resolved in the enlarged central region. Based on visual inspection, the smallest resolvable feature was identified as Group 8, Element 3 of the USAF 1951 target, where the line pairs remained di.......

Discussion

This DIY platform (Figure 10) integrates a compact optical layout, programmable illumination, and open-source control software to provide an accessible solution for long-term imaging inside standard incubators. Compared with commercial time-lapse incubators and integrated imaging systems^7,8, this 3D-printed platform substantially reduces cost while maintaining the resolution required for routine observation of cultured cells and earl.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
10× LWD Infinity Objective	Standard Part	https://item.taobao.com/item.htm?id=750667327346	Infinity-corrected long-working-distance objective, 10×
2 × 20 Pin Male Header	Standard Part	–	2.54 mm pitch GPIO header; quantity: 1
5 mm ID Ball Bearing	Standard Part	–	φ5 × 10 × 5 mm ball bearing; quantity: 1
Aviation Connectors	Standard Part	–	2-pin and 3-pin aviation plugs; quantity: 2
Camera Module (IMX219)	Raspberry Pi	IMX219	Raspberry Pi Camera V2; quantity: 1
CSI cable holder A	Custom	Custom	3D-printed component: 3D-Print - csi_cable_holder_A.STL
CSI cable holder B	Custom	Custom	3D-printed component: 3D-Print - csi_cable_holder_B.STL
Diffuse Acrylic Plate	Standard Part	–	72 × 107 mm, 1 mm thick; quantity: 1
Enclosure lower	Custom	Custom	3D-printed component: 3D-Print - enclosure_lower.STL
Enclosure upper	Custom	Custom	3D-printed component: 3D-Print - enclosure_upper.STL
General well-plate holder	Custom	Custom	3D-printed component: 3D-Print - general_well-plate_holder.STL
High-Transmission Acrylic Plate	Standard Part	–	116 × 194 mm, 2 mm thick; quantity: 1
Light shell A	Custom	Custom	3D-printed component: 3D-Print - light_shell_A.STL
Light shell B	Custom	Custom	3D-printed component: 3D-Print - light_shell_B.STL
Linear Bearing LM5UU	Standard Part	LM5UU	φ5 × 10 × 15 mm linear bearing; quantity: 4
M2 Countersunk Self-Tapping Screw	Standard Part	–	M2 × 8 mm; quantity: 4
M2 Self-Tapping Button-Head Screw	Standard Part	–	M2 × 10 mm; quantity: 8
M2.5 Hex Socket Screw (10 mm)	Standard Part	–	M2.5 × 10 mm; quantity: 12
M2.5 Hex Socket Screw (8 mm)	Standard Part	–	M2.5 × 8 mm; quantity: 12
M3 Button-Head Screw	Standard Part	–	M3 × 20 mm; quantity: 4
M3 Button-Head Screw + M3 Nut Set (10 mm)	Standard Part	–	M3 × 10 mm screw + M3 nut; quantity: 20 sets
M3 Button-Head Screw + M3 Nut Set (12 mm)	Standard Part	–	M3 × 12 mm screw + M3 nut; quantity: 20 sets
M3 Button-Head Screw + M3 Nut Set (15 mm)	Standard Part	–	M3 × 15 mm screw + M3 nut; quantity: 20 sets
Mainboard bracket	Custom	Custom	3D-printed component: 3D-Print - mainboard_bracket.STL
microSD Card	Standard Part	–	minimum 8 GB; 32 GB or larger recommended)
Middle frame	Custom	Custom	3D-printed component: 3D-Print - middle_frame.STL
OpenLapse Controller Board	Custom	Custom	PCB files available in Supplemental Files
OpenLapse Illumination Array Board	Custom	Custom	PCB files available in Supplemental Files
Raspberry Pi Camera FPC Cable	Standard Part	https://item.taobao.com/item.htm?id=774536454547	22-pin to 15-pin, 40 mm CSI cable
Raspberry Pi Zero 2 W	Raspberry Pi	–	Main control computer; quantity: 1
RMS-to-SML Adapter Ring	Standard Part	https://item.taobao.com/item.htm?id=711654577589	RMS to SML conversion ring
SML bracket	Custom	Custom	3D-printed component: 3D-Print - SML_bracket.STL
SML05 Tube Holder	Standard Part	SML05 (https://detail.tmall.com/item.htm?id=613460973426)	Optical holder
SMLRR Retaining Ring	Standard Part	SMLRR (https://detail.tmall.com/item.htm?id=613460973426)	Retaining ring
T5 Cut-Flange Lead Screw Nut (2 mm pitch)	Standard Part	–	T5 × 2 mm pitch lead screw nut; quantity: 2
T5 Lead Screw Nut (2 mm pitch)	Standard Part	–	T5 × 2 mm pitch lead screw nut; quantity: 1
TMC2209 Driver Module	MKS	TMC2209 (https://item.taobao.com/item.htm?id=782341270726)	Stepper motor driver
Tube Lens	Standard Part	https://item.taobao.com/item.htm?id=590466758183	φ25.4 mm, focal length 50 mm
X-axis bracket	Custom	Custom	3D-printed component: 3D-Print - x-axis_bracket.STL
X-Axis Lead Screw Stepper Motor	Standard Part	https://item.taobao.com/item.htm?id=922508067421	28-series motor with T5 × 200 mm lead screw, 2 mm pitch
X-Axis Linear Rail with Carriage	Standard Part	https://item.taobao.com/item.htm?id=837345295063	MGW9-200 mm rail with MGW9H carriage
XH 2.54 mm 3-Pin Cable	Standard Part	–	Male-to-male connector cable, 400 mm; quantity: 1
Y-axis bracket A	Custom	Custom	3D-printed component: 3D-Print - y-axis_bracket_A.STL
Y-axis bracket B	Custom	Custom	3D-printed component: 3D-Print - y-axis_bracket_B.STL
Y-Axis Guide Rod	Standard Part	–	φ5 × 170 mm precision shaft; quantity: 1
Y-Axis Lead Screw Stepper Motor	Standard Part	https://item.taobao.com/item.htm?id=788375595753	28-series motor with T5 × 150 mm lead screw, 2 mm pitch
Z-axis bracket A	Custom	Custom	3D-printed component: 3D-Print - z-axis_bracket_A.STL
Z-axis bracket B	Custom	Custom	3D-printed component: 3D-Print - z-axis_bracket_B.STL
Z-Axis Guide Rod	Standard Part	–	φ5 × 80 mm precision shaft; quantity: 1
Z-Axis Lead Screw Stepper Motor	Standard Part	https://item.taobao.com/item.htm?id=922508067421	28-series motor with T5 × 50 mm lead screw, 2 mm pitch