Interlab

Introduction

Peking 2018 joined the fifth Interlab measurement study. This year we helped to answer this question: Can we reduce lab-to-lab variability in fluorescence measurements by normalizing to absolute cell count or colony-forming units (CFUs) instead of OD?

The introduction of this year Interlab: https://2018.igem.org/Measurement/InterLab

Equipment Information

Plate Reader: Perkin Elemer EnSpire ^TM Multilabel Reader 2300

Flow Cytometry: BD LSRFortessa ^TM Cell Analyzer

96 - Well Pate: Corning Incorporated Costar^®️ 3603

Results

1.

Calibration 1: OD600 Reference point - LUDOX

Figure. 1 The result of LUDOX calibration. The correction factor of our plate reader is 3.316

2.

Calibration 2: Particle Standard Curve – Microsphere

Figure. 2 The result of particle calibration.


(a) Particle Standard Curve - Linear	(b) Particle Standard Curve - Log Scale
Figure. 3 The result of particle standard curve

3.

Calibration 3: Fluorescence standard curve – Fluorescein

Figure. 4 The result of fluorescein calibration.


(a) Fluorescence Standard Curve - Linear	(b) Fluorescence Standard Curve - Log Scale
Figure. 5 The result of fluorescence standard curve

Cell Measurement

Materials:

Competent cells (Escherichia coli strain DH5α)
LB (Luria Bertani) media
Chloramphenicol (stock concentration 25 mg/mL dissolved in EtOH)
50 ml Falcon tube (or equivalent, preferably amber or covered in foil to block light) Incubator at 37°C
1.5 ml eppendorf tubes for sample storage
Ice bucket with ice
Micropipettes and tips
96 well plate, black with clear flat bottom preferred