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28.1.2 Indirect Enzyme-Linked Bioassays on 96-Microwell
Format Plate
Indirect enzyme-linked bioassay of Ag-Ab was used as a reference test methods for
96-microwell formats
28.1.3 SSHG Spectroscopy Sampling and Analysis Procedure
for 96-Microwell Format
The aim of this stage of the study is reproducing several replicates of the optimised
immunoassays. The objective of this stage is to investigate the application of SSHG
spectroscopy on biomolecular interactions by comparing SSHG modulation to the
reference method readings.
The following SSHG readings were recorded during this phase of study as
follows:
1. From empty surfaces, i.e. prior to depositing Ag, denoted as SSHG e .
2. After depositing Ag and washing excessive deposited biomolecules and drying
phase, denoted as SSHG wd .
3. After forming Ag-Ab complex, denoted as SSHG pb .
SSHG Readings
The general procedure for SSHG spectroscopy on immunoassays and DNA bioassays is summarised as follows:
1. Prepare replicates of the optimised enzyme-linked bioassay following the procedure described. NB. Only dilution steps for which the best linear fit is observed
are needed to prepare these replicates in this step.
2. Prior to step 1, programme the scanner to select five random selected spots from
each microwell and record SSHG e (Namely, the centre of each microwell and
other four radial points located at 0.25 mm from the centre were selected). At
step 2 of the preparation procedure, programme the scanner to scan the spots
selected in 2 and record SSHG wd .
3. At step 5 of the preparation procedure, programme the scanner to scan the spots
selected in 2 and record SSHG pb values.
Analysis of SSHG Readings
1. Compute SSHG modulation due to Ag deposition, denoted as
∆SSHG depos-empty = SSHG wd -SSHG e .
2. For ∆SSHG depos-empty compute the following statistical parameters: mean, median,
standard deviation, variance and test the data for normal distribution using parametric tests. These parameters are used to examine central tendency and variance
of data.
A. M. Soliman
28.1.2 Indirect Enzyme-Linked Bioassays on 96-Microwell
Format Plate
Indirect enzyme-linked bioassay of Ag-Ab was used as a reference test methods for
96-microwell formats
28.1.3 SSHG Spectroscopy Sampling and Analysis Procedure
for 96-Microwell Format
The aim of this stage of the study is reproducing several replicates of the optimised
immunoassays. The objective of this stage is to investigate the application of SSHG
spectroscopy on biomolecular interactions by comparing SSHG modulation to the
reference method readings.
The following SSHG readings were recorded during this phase of study as
follows:
1. From empty surfaces, i.e. prior to depositing Ag, denoted as SSHG e .
2. After depositing Ag and washing excessive deposited biomolecules and drying
phase, denoted as SSHG wd .
3. After forming Ag-Ab complex, denoted as SSHG pb .
SSHG Readings
The general procedure for SSHG spectroscopy on immunoassays and DNA bioassays is summarised as follows:
1. Prepare replicates of the optimised enzyme-linked bioassay following the procedure described. NB. Only dilution steps for which the best linear fit is observed
are needed to prepare these replicates in this step.
2. Prior to step 1, programme the scanner to select five random selected spots from
each microwell and record SSHG e (Namely, the centre of each microwell and
other four radial points located at 0.25 mm from the centre were selected). At
step 2 of the preparation procedure, programme the scanner to scan the spots
selected in 2 and record SSHG wd .
3. At step 5 of the preparation procedure, programme the scanner to scan the spots
selected in 2 and record SSHG pb values.
Analysis of SSHG Readings
1. Compute SSHG modulation due to Ag deposition, denoted as
∆SSHG depos-empty = SSHG wd -SSHG e .
2. For ∆SSHG depos-empty compute the following statistical parameters: mean, median,
standard deviation, variance and test the data for normal distribution using parametric tests. These parameters are used to examine central tendency and variance
of data.
A. M. Soliman