Drug Dosing Calculator User Guide

Chapter 1. Overview

1.1. Drug Dosing Calculator Overview

Axion’s Drug Dosing Calculator calculates the concentrations and volumes needed to prepare dosing solutions for plate-based experiments.

1.2. Stock Solution Conditions

1. Enter your concentrated stock solution (Stock) and the vehicle concentration in the stock solution (Vehicle).
   1. 1.1 Ensure proper units are selected using the dropdown option.
   2. 1.2 For example, a compound prepared in all DMSO would typically need to be entered as 100% DMSO.

1.3. Target Dose Conditions

1. Enter your target cell plating density in the Cells per Well field, as well as the cell plating volume (V_plating) and number of wells to be plated.
2. Multiple plating densities may be added or removed with the plus and minus buttons.
   1. 2.1 Note: Order the list of dosage concentrations with proper units from highest to lowest.
   2. 2.2 Additionally, Spare Volume provides room to account for extra solution in each aliquot due to pipetting loss or dead volume.

1.4. Well Conditions

1. Enter your well-starting solution volume (Starting Well Volume) and the % of desired final vehicle concentration into Final [Vehicle].
   1. 1.1.Dose Volume represents the volume of drug solution added to the well.
2. To add to the initial volume or replace an equivalent amount of volume with dose solution, click on Add To or Replace respectively.

1.5. Solution Results

Then, click Calculate Results. The following outputs are provided:

1. Product identifies the solution being prepared.
2. [Conc] provides the target concentration of the product.
3. Solution identifies the source solution used to prepare the product.
4. V_solution provides the required volume of the source solution.
   1. 4.1 It is rounded to the nearest 0.1 µL and is not allowed to be less than 10 µL. V_diluent is adjusted accordingly.
5. Diluent identifies the diluent used to prepare the product.
6. V_diluent provides the required volume of diluent.

For each solution (Product) you need to make, add Vdiluent to V_solution slowly, mixing as needed.

• Note: Adding V_solution to V_diluent may cause solubility issues. Always dilute high to low.
• Note: The starting stock or vehicle concentration is considered too low if the desired final vehicle concentration cannot be achieved.

Chapter 2. Definitions

2.1. Starting Conditions

Stock Solution Conditions

2.2. Target Conditions

Target Dose Conditions

2.3. Well Conditions

Experiment Setup

2.4.Results

Results

Chapter 3. Equations

3.1. Solution Design

Solution Design Calculations:
(n = number of drug solutions or length of the array)

Volume Calculations:

For working solutions:

For intermediates:

Vehicle & Diluent Preparation Calculations: