Compound Administration Planning

COMPOUND ADMINISTRATION PLANNING

1Experiment mode: choose dosing design, administration method, final-volume behavior, and solvent limit.
2Workflow-specific volumes: enter the starting well volume, any concentration already present at time 0, and the addition or cast-return volume.
3Spatial distribution and doses: define conditions, compound names, concentrations or cumulative dose series, and assign them to chips or wells.
4Dose-dilution table: enter the final concentrations to reach for each compound and add rows if needed.
5Dilution and solution preparation: adjust compound-specific stocks if needed, then follow the generated stock, intermediate, powder, and per-condition instructions.
5.1Compound-specific main and intermediate stocks: enter or adjust the main-stock concentration for each compound; intermediate stock(s) are calculated automatically.
5.2Main stock from powder: use only when a compound stock is prepared from weighed powder.
5.3Preparation strategy and condition instructions: choose separate or serial dilution and follow the generated condition instructions.
6Print or save workflow: keep the experimental layout and dilution plan available at the hood, or save it for later import.

1Experiment mode

Dosing design
Administration method
Maintain constant final well volume?
Maximum final solvent v/v % allowed in the well

Vehicle and solvent compatibility guide

Use the lowest vehicle concentration that still keeps the compound soluble and pipettable. Match the vehicle concentration in control wells and validate any non-standard vehicle for the specific cell type, exposure duration, and readout.

Vehicle / excipient	Recommended final limit in well	Practical guidance
Culture medium / recording medium	Preferred whenever possible	Best choice for working dilutions because it preserves osmolarity, salt balance, pH, nutrients, and cell compatibility.
Water / sterile ultrapure water	≤0.5–1% v/v preferred; avoid >2–3% unless validated	Usually less biologically active than organic solvents, but can still alter osmolarity, pH, salts, nutrients, and final volume. Dilute stocks into medium before dosing when possible.
DMSO	≤0.05% v/v preferred; 0.1% only if validated	Common default for hydrophobic compounds, but neuronal activity can be sensitive even at low levels. Always include matched vehicle controls.
Ethanol	0.01–0.05% v/v; avoid >0.1%	Can directly modulate neuronal activity. Keep the final concentration as low as possible and validate with vehicle controls.
Methanol	0.01–0.05% v/v; avoid when possible	Not ideal for live neuronal cultures. Use only when necessary and after compatibility testing.
PEG 400	0.05–0.1% v/v	Useful co-solvent for some compounds; monitor osmolarity, viscosity, and precipitation after dilution into medium.
Propylene glycol	0.05–0.1% v/v	Possible co-solvent. Validate carefully because it may alter neuronal activity or medium properties.
Glycerol	0.05–0.1% v/v	Limited usefulness for hydrophobic compounds and may affect osmolarity and viscosity.
HP-β-cyclodextrin	~0.01–0.1% w/v	Aqueous solubilizer rather than a classical solvent. Validate separately and use matched controls.
Tween-80 / Polysorbate 80	0.001–0.01% v/v	Surfactant with possible membrane and activity effects. Use only if required for solubility.
Cremophor EL / Kolliphor	0.001–0.01% v/v	Strong solubilizer with high artifact risk in live-cell assays. Avoid unless necessary and validated.
DMF	≤0.01% v/v; avoid >0.05%	Strong solvent and not a neutral vehicle for live neurons. Use only with careful validation.
NMP / Pharmasolve	≤0.01% v/v; generally avoid	Biologically active vehicle and generally a poor choice for neuronal assays.
Acetonitrile	≤0.01% v/v; avoid for live cultures	More appropriate for analytical chemistry than live-cell dosing.

Example concentrations from user publications

Use these as starting examples only. Always adapt the final concentration range to the specific cell type, preparation, exposure duration, endpoint, and solvent tolerance. Vehicle controls should normally match the highest final solvent percentage used in the experiment, especially for DMSO or other non-water vehicles.

Compound	Sample	Concentration	Expected general effect / mechanism	Ref
Diazepam	Sliced hiPSC-derived brain organoids	3–50 µM	Benzodiazepine positive allosteric modulator of GABA_A receptors; expected to strengthen inhibitory signaling and often reduce excitability.	1
Picrotoxin (PIC)	hiPSC-derived neurons hiPSC-derived neural spheroids	0.1–10 µM 0.3–30 µM	GABA_A receptor channel blocker/antagonist; expected to remove inhibitory tone and increase firing, bursting, or synchrony.	2
CNQX	hiPSC-derived cortical organoids hiPSC-derived neural spheroids	20 µM 0.3–30 µM	AMPA/kainate receptor antagonist; expected to reduce fast glutamatergic excitation.	2
Bicuculline (BIC)	Sliced hiPSC-derived brain organoids	20 µM	Competitive GABA_A receptor antagonist; expected to disinhibit networks and increase activity.	3
Tetrodotoxin (TTX)	Sliced hiPSC-derived brain organoids	1 µM	Voltage-gated sodium channel blocker; expected to suppress action potential generation and network activity.	3
NBQX	Sliced hiPSC-derived brain organoids iNGN-EGFP hiPSC-derived neurons	10 µM 10 µM	AMPA/kainate receptor antagonist; expected to reduce excitatory synaptic drive.	1, 4
Gabazine (GBZ)	Sliced hiPSC-derived brain organoids iNGN-EGFP hiPSC-derived neurons	10 µM 10 µM	Selective competitive GABA_A receptor antagonist; expected to disinhibit networks.	1, 4
Muscimol	hiPSC-derived neurons	10 µM	GABA_A receptor agonist; expected to enhance inhibition and suppress firing/bursting.	6
AP5	TF-induced iPSC neurons	0.1–10 µM	NMDA receptor antagonist; expected to reduce NMDA-dependent excitation/plasticity.	5
4AP	TF-induced iPSC neurons	10–300 µM	Voltage-gated potassium channel blocker; prolongs action potentials and increases neuronal excitability, firing, and network bursting.	5
KCl	Primary rat vagal afferent neurons	20 mM	At high concentration (e.g. 20 mM), strong depolarizing perturbation that can silence activity; at lower concentrations may acutely depolarize neurons.	7

Reference list: 1 Sharf et al.; 2 Han et al.; 3 Schröter et al.; 4 Habibey et al.; 5 Lin et al.; 6 Kelley et al.; 7 Girardi et al..

2Workflow-specific volume inputs: Direct addition

Direct-addition setup

Initial well volume (µL)
Initial concentration (µM)
Direct-addition aliquot (µL)

Direct-addition volume suggestion

Prefer small added volumes, typically 5-30 µL, to limit perturbation of the sample secretome and to avoid introducing a medium-change effect. Use the smallest practical volume that still allows accurate pipetting and feasible stock dilution. After addition, gently mix in the well to reduce local concentration gradients; keep the mixing method consistent across compound and vehicle wells.

3Experiment batch and spatial distribution

Number of conditions to be tested
Spatial distribution

Vehicle and solvent compatibility guide

Example concentrations from user publications

Direct-addition volume suggestion

Cast-and-return instruction

Working-solution dilution strategy

Initial well volume (µL)
Initial concentration (µM)
Casted medium volume (µL)