Fully automated illumination study series for modern cancer drug development

Modern Cancer Drug Development

Fully automated illumination study series for modern cancer drug development

In vitro cell viability and dose escalation study for developing photosensitive or photoactivated drugs using Modulight ML7710 medical laser and ML8500 automated illumination system.

So how was it done?

 

STUDY PLAN

The goal was to investigate the effect of irradiance and light dose on a cancer cell line while keeping the photosensitive drug dosing constant. The well-by-well dose escalation plan was done in tabular format.

target

TARGET

Cancer cell line incubated at 37°C in acidic culture

drug-type

DRUG TYPE

Photosensitive or photoactivated cancer drug (constant)

illumination

ILLUMINATION

Variable dose of
0/10/20/25/50/75/100 J/cm2

irradiance

IRRADIANCE

100 and 600 mW/cm2

 

Standard 96-well microplate

A standard 96-well microplate was divided so that half of the wells were illuminated with irradiance of 600 mW/cm2 (columns 1-6), and the other half with 100 mW/cm2 (columns 7-12).

The dose was increased by the row, using doses of 0 – 5 – 10 – 25 – 50 – 75 – 100 J/cm2.

This way, there were 6 technical replicates of each combination of dose & irradiance, which allows calculation of standard deviation for each data point.

 

Dose (J/cm2) / Irradiance (mW/cm2)

1 2 3 4 5 6 7 8 9 10 11 12
A 0
600
0
600
0
600
0
600
0
600
0
600
0
100
0
100
0
100
0
100
0
100
0
100
B 5
600
5
600
5
600
5
600
5
600
5
600
5
100
5
100
5
100
5
100
5
100
5
100
C 10
600
10
600
10
600
10
600
10
600
10
600
10
100
10
100
10
100
10
100
10
100
10
100
D 15
600
15
600
15
600
15
600
15
600
15
600
15
100
15
100
15
100
15
100
15
100
15
100
E 25
600
25
600
25
600
25
600
25
600
25
600
25
100
25
100
25
100
25
100
25
100
25
100
F 50
600
50
600
50
600
50
600
50
600
50
600
50
100
50
100
50
100
50
100
50
100
50
100
G 75
600
75
600
75
600
75
600
75
600
75
600
75
100
75
100
75
100
75
100
75
100
75
100
H 100
600
100
600
100
600
100
600
100
600
100
600
100
100
100
100
100
100
100
100
100
100
100
100

The dose is displayed on the first row and the irradiance on the second row.

 

Experimental set up

modulight-ml8500

Well plate illumination

Modulight ML8500 automated illumination system

Read more

modulight-ml7710

Light source

Modulight ML7710 multi-wavelenght medical laser system

Read more

cellometer-auto2000

Cell counting

Nexcelom Cellometer Auto2000 cell viability counter

STUDY EXECUTION

The study of photosensitive or photoactivated drugs requires a controlled and systematic sample illumination process. Modulight ML8500 automated illumination system and ML7710 medical laser ensure the repeatable and accurate illumination of each sample. ML7710 supports up to 8 wavelengths allowing research of multiple drugs, or combining fluorescence measurements to the study plan.

Extracting and counting cells

Extracting and counting the cells with Cellometer Auto2000 (unit / ml)

Diluting the cells to 1 million / ml

Centrifuging and separating the cancer cells

Centrifuging and separating the cancer cells with cell stripper

Adding saline based new medium and serum

Adding saline based new medium and serum

Mixing and incubating the sample

Mixing and incubating the sample

Flushing cells free from free-floating drug

Washing the cells from free-floating drug

Confirming the drug bindings

Confirming the drug bindings

Dispensing the cells to be radiated

Dispensing the cells to be  radiated on 96-well sample plate

Illuminating the L96 with Modulight

Illuminating the 96-well sample plate with Modulight ML8500 and ML7710

Incubating to eliminate close-to-death cells

Incubating to eliminate close-to-death cells for appr. 1.5 h

result

ML8500 automated illumination system enables the researchers to investigate the effect of multiple independently variable parameters easily and reliably.

  1. Increase in dose (J/cm2) and in the overall cell death reached saturation at 50 J/cm2
  2. The efficacy of the drug was dependent on the irradiance. High irradiance (600 mW/cm2) resulted in decreased killing ratio at lower light dose range, which is supported by the literature and suspected to be caused by oxygen depletion in the sample [1] or less effective exhaustion of cell antioxidant responses [2].
  3. ML8500 automated illumination system enabled variation of 2 independent parameters (dose and irradiance) over 96-well plate in a single experiment.
100 mW/cm2 600 mW/cm2
Log LLD50 0.9908 1.317
HillSlope -2.431 -4.106
LLD50 9.79 20.75

 


fully-automated-testing

Fully automated testing

better-validability

Better validability and reliability

more-tolerable-treatment

Optimized drug & light dose for the patient


 

 

 

 

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