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KEEP RED LINE OFF SLIDE Corporate Overview November 2025 ©2025 Pulse Biosciences, Inc. All rights reserved. 1
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KEEP RED LINE OFF SLIDE All statements in this presentation that are not historical are forward-looking statements, including, among other things, statements concerning the effectiveness of the Company’s nPulse technology to non-thermally clear cells while sparing adjacent noncellular tissue, statements concerning the potential market opportunities for the use of nsPFA energy to treat atrial fibrillation or benign thyroid nodules and whether these markets might grow in the future; statements concerning early clinical successes and whether they are predictive of the safety and effectiveness of any medical device, statements concerning the Company’s future clinical and regulatory initiatives anywhere in the world, such as whether the FDA will approve the Company’s presently pending IDE submission, the Company’s expectations, whether stated or implied, regarding whether the Company’s nsPFA technology will become a disruptive, superior and durable treatment option for treating atrial fibrillation or any other medical condition, and other future events. These statements are not historical facts but rather are based on Pulse Biosciences’ current expectations, estimates, and projections regarding Pulse Biosciences’ business, operations and other similar or related factors. Words such as “may,” “will,” “could,” “would,” “should,” “anticipate,” “predict,” “potential,” “continue,” “expects,” “intends,” “plans,” “projects,” “believes,” “estimates,” and other similar or related expressions are used to identify these forward-looking statements, although not all forward-looking statements contain these words. You should not place undue reliance on forward-looking statements because they involve known and unknown risks, uncertainties, and assumptions that are difficult or impossible to predict and, in some cases, beyond Pulse Biosciences’ control. Actual results may differ materially from those in the forward-looking statements as a result of a number of factors, including those described in Pulse Biosciences’ filings with the Securities and Exchange Commission. Pulse Biosciences undertakes no obligation to revise or update information in this release to reflect events or circumstances in the future, even if new information becomes available. This presentation and any documents incorporated by reference may contain market data that we obtain from industry sources. These sources do not guarantee the accuracy or completeness of the information. Although we believe that our industry sources are reliable, we do not independently verify the information. The market data may also include projections that are based on other projections. While we believe these assumptions and projections are reasonable and sound, as of the date hereof, actual results may differ from these projections. Forward Looking Statements ©2025 Pulse Biosciences, Inc. All rights reserved. 2
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KEEP RED LINE OFF SLIDE ©2025 Pulse Biosciences, Inc. All rights reserved. Our Mission To build a thriving viable company by providing revolutionary and life-changing therapy for multiple diseases with next-generation Nanosecond Pulsed Field technology. ©2025 Pulse Biosciences, Inc. All rights reserved. 3
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Inventing and harnessing nPulse technology dates back two decades • Differentiated approach focused on novel therapy development • 10+ years in development at Pulse Biosciences • $400mm Development Investment to date • >1,100 industry-wide publications to date • Multiple FDA Clearances • Breakthrough Designation +100 Pending patent applications +215 Issued patents globally owned & licensed Patent Portfolio 2025 History of Pioneering Nanosecond PFA Development ©2025 Pulse Biosciences, Inc. All rights reserved. Creating wide and deep IP portfolio covering nPulse energy and system Continued development and patent filings covering systems, applications, and methods Pulse Biosciences is the leader in designing and engineering nPulseTM technology 4
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Experienced Technologists, Operators and Clinicians Form Proven Leadership Team Established Board of Directors Robert (Bob) W. Duggan Co-Chairman of the Board of Directors Mahkam "Maky" Zanganeh, DDS Director Manmeet S. Soni Director Richard van den Broek Director Dr. Gan Dunnington Chief Medical Officer, Cardiac Surgery Dr. Niv Ad Chief Science Officer, Cardiac Surgery Dr. David Kenigsberg Chief Medical Officer, Electrophysiology ©2025 Pulse Biosciences, Inc. All rights reserved. Darrin Uecker Chief Technology Officer Director Kevin Danahy Chief Commercial Officer Mitch Levinson Chief Strategy Officer Paul LaViolette Chief Executive Officer Co-Chairman of the Board 5 Jon Skinner Chief Financial Officer
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Financial Snapshot Strong balance sheet as of 9/30/2025 • Cash and cash equivalents balance of $95.2 mm • No debt • Cash burn of approximately $36mm in 2024 • Cash burn increasing in 2025 to support pilot program expansion and multiple IDEs ($13.0mm of cash used in operating activities in 3Q25) Generated initial revenue from Vybrance limited market release • 3Q25 revenue totaled $86k +80% Insider Ownership ©2025 Pulse Biosciences, Inc. All rights reserved. 6
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Unique Mechanism of Action of nsPFA ©2025 Pulse Biosciences, Inc. All rights reserved. 7 Stimulates elegant, precise Regulated Cell Death (RCD) in cells without thermal spread Nanosecond pulsed electrical fields delivered to cells Higher amplitude energy pulses penetrate the cell membrane and disrupt internal cellular function in organelles, leading to regulated cell death vs. inflammatory response caused by thermal / microPFA Unlike thermal and microPFA modalities, nsPFA directly impacts cellular structures while sparing noncellular tissue (primarily collagen)
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Pulse Biosciences Difference: Nanosecond PFA Delivers Higher Amplitude Shorter Duration Pulses with Significant Advantages Versus Microsecond PFA nPulse technology’s novel mechanism activates the body’s natural healing process ©2025 Pulse Biosciences, Inc. All rights reserved. 8 Clinical benefits – safety and efficacy ✓ Nonthermal: no thermal damage or spread beyond the electrical field ✓ Lower total energy delivered due to ultrashort pulse durations, despite higher pulse amplitude ✓ Faster deep ablations with single energy delivery ✓ Selectively treats only organelle-bearing structures, spares nerves, vessels, structural tissue ✓ Reduces stimulation to nerves and muscle ✓ Much lower energy reduces electrode design constraints of standard PFA
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Pulse Biosciences’ nPulse Platform Unlocks >$6B in first 3 target markets to >$16B global including Cardiac Surgery EP Catheter $3.0B 8 $1.8B 6,7 Thyroid Application in Soft Tissue Ablation $1.3B 1,2,3,4,5* Pulse will create, expand, and upend these respective markets with therapy innovation, market penetration and global expansion ©2025 Pulse Biosciences, Inc. All rights reserved. $2.8B 1,2,3,4,5* $6.0B 6,7 $8.0B 9 *based on reimbursement updates and asp assumptions 9 U.S. Addressable Annual Market Global Potential Market
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FACTORS INFLUENCING THE STRATEGY INCLUDE: • New (thyroid) or established (open/EP) market • Presence of direct competition • Efficiency of launch investment • Scale and simplicity of platforms • Therapy synergies • Enabling differentiation for a strategic partner >$6B combined Market Entry ©2025 Pulse Biosciences, Inc. All rights reserved. Segment business models will be determined based on efficiency to create, expand and upend respective markets 10 Expedite time to access patients Go direct Via Partnership
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nPulse Vybrance Percutaneous Electrode: Create an Untapped Market in Thyroid Opening a new era in therapy for soft tissue ablation such as benign thyroid nodules Thyroid Therapy | Surgical Ablation | AF Ablation ©2025 Pulse Biosciences, Inc. All rights reserved. March 2024 11
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Benign Thyroid Market Opportunity ©2025 Pulse Biosciences, Inc. All rights reserved. Drivers for Pulse in the Benign Thyroid Market • Thyroid-sparing procedure • Nonsurgical intervention • Potential to eliminate up to ~150k thyroidectomies yearly ~100k Enter “Watchful Waiting” ~150k Benign thyroidectomies BTN patients diagnosed in U.S. ~250k3,4,5 $1B+ Annual U.S. TAM nPulse technology is a new paradigm for surgical patients and watchful waiters ©2025 Pulse Biosciences, Inc. All rights reserved. Thyroid Therapy | Surgical Ablation | AF Ablation Newly created untapped market 12
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Create an Untapped Market in Thyroid nPulse therapy provides significant clinical benefits for physicians and patients ©2025 Pulse Biosciences, Inc. All rights reserved. Thyroid Therapy | Surgical Ablation | AF Ablation • Safe: Spares nerves, vessels, critical structures • Nonthermal - does not cause fibrosis or RF scar ball formation • Significant volume reduction and symptomatic relief at 1 month • High patient satisfaction – Improved cosmesis – Soft, natural treatment area Baseline 1 month post 12 months post Prof. Stefano Spiezia – ATA 2024 Presentation 13
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Thyroid Market Development Status Generated initial revenue through commencement of nPulse Vybrance limited market release in 3Q25 ©2025 Pulse Biosciences, Inc. All rights reserved. • Soft Tissue Ablation – FDA 510(k) Cleared • Initiated a multi-center clinical study, PRECISE-BTN, for the treatment of benign thyroid nodules (BTN) • Enrolled over 40% of study subjects • Increasing procedure volumes • Over 200 patients treated across the pilot program, the PRECISE-BTN Study, and initial commercial procedures with positive results • Geographic commercial concentration 14 Thyroid Therapy | Surgical Ablation | AF Ablation
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nPulse Cardiac Clamp: Expanding Usage in the Surgical Cardiac Ablation Market nPulse therapy for Surgical AF ©2025 Pulse Biosciences, Inc. All rights reserved. Thyroid Therapy | Surgical Ablation | AF Ablation 15
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Surgical Cardiac Ablation Market Opportunity ©2025 Pulse Biosciences, Inc. All rights reserved. Thyroid Therapy | Surgical Ablation | AF Ablation Drivers for Pulse in the Surgical Cardiac Ablation Market • 80,000+ U.S. patients per year with pre-operative AF • Only ~30% being treated with RF technology • Pulse expects to be first to market with a PFA solution • RF technology: safety and efficacy concerns nPulse technology expands the market Annual open-heart procedures in U.S. 300K11 216k Annual U.S. Patients 72% Patients at high risk of developing post-op AF13 28% Patients have pre-operative AF12 Future Potential Prophylactic AF 16
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Expanding Usage in the Surgical Cardiac Ablation Market Speed and Versatility for Cardiac Surgical Applications • Speed, ease of use and safety of nPulse therapy address key physician concerns limiting adoption of thermal modalities – Capable of transmural ablations in seconds – Fully automated ablation independent of tissue thickness or type – Nonthermal, eliminating risk of damage to surrounding critical structures, e.g. esophagus ©2025 Pulse Biosciences, Inc. All rights reserved. Bipolar Clamp Thyroid Therapy | Surgical Ablation | AF Ablation 17
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Surgical Cardiac Ablation Status • Received FDA IDE approval and enrolled the first subject in the nPulse Cardiac Surgery System Pivotal Study (NANOCLAMP AF) • Treated 44 patients as part of the multi-center feasibility study underway in Europe • Late-breaking data from the European feasibility study, presented at the 39th EACTS Annual Meeting, showed durable, consistent pulmonary vein and posterior box isolation achieved in the first 24 patients FDA approved IDE pivotal study underway ©2025 Pulse Biosciences, Inc. All rights reserved. Thyroid Therapy | Surgical Ablation | AF Ablation 18
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nPulse Cardiac Catheter: Upending AF Ablation in the EP Market nPulse therapy for AF ©2025 Pulse Biosciences, Inc. All rights reserved. Thyroid Therapy | Surgical Ablation | AF Ablation 19
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EP Market Opportunity AF Ablation ©2025 Pulse Biosciences, Inc. All rights reserved. ©2025 Pulse Biosciences, Inc. All rights reserved. Thyroid Therapy | Surgical Ablation | AF Ablation Market growing at 10–15% CAGR 14 ~1.9M 15 U.S. patients diagnosed with AF annually Global Atrial Fibrillation (AF) Disease State: >$8B 9 Electrophysiology Market Drivers for Pulse in the EP AF Ablation Market • Drop-in workflow replacement • Speed = Improved efficiency over 1st gen PFA devices • Less configurations, rotations, applications • Enhanced lesion quality through nPulse therapy nPulse technology upends the EP market with a novel and differentiated energy 20
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Customized nPulse Electrode Design nPulse Catheter for Pulmonary Vein Isolation Achieves circumferential lesions with continuous ring electrodes for “single-shot” Pulmonary Vein Isolation (PVI) ablation • 5 seconds ablation cycle • <10 minutes ablation time with moderate experience • Single circumferential lesion—no rotation • Deeper lesions and low neuromuscular stimulation • Nonthermal isolation of vein • Design allows for versatility in left atrial utilization • Sensing electrodes and magnetic sensor for EAM integration 2-Day Endocardial Surface ~5cm Diameter (Porcine) ©2025 Pulse Biosciences, Inc. All rights reserved. Thyroid Therapy | Surgical Ablation | AF Ablation 21
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# Configurations Energy deliveries per target Micro-PFA Compared to nPulse for Electrophysiology Current workflow and case time 8 2 ©2025 Pulse Biosciences, Inc. All rights reserved. Device rotations to “connect the dots”R E Q U I R E D N O N E R E Q U I R E D Patient paralyticsR E Q U I R E D N O N E R E Q U I R E D Pulse durationMicrosecond Nanosecond 1 2 Average ablation time30 minutes16 10 minutes Lesion quality/depth4–6mm17 6–9mm18 22
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Initial 360 Cardiac Catheter Feasibility Study Data ©2025 Pulse Biosciences, Inc. All rights reserved. Dr. Johan Vijgen Jessa Hasselt Hospital Post map after Jessa Hasselt’s first case Thyroid Therapy | Surgical Ablation | AF Ablation 23 Late-breaking data from the first 30 patients treated demonstrated exceptional results, as presented by Vivek Reddy, MD, at the AF Symposium in January 2025 • All (100%) lesions were acutely successful with conduction block • Success rate of PVI at ~3 months was 92.4% (109/118) • Total PVI ablation time was 11.6±4.5 minutes • Total procedure and fluoroscopy times were 88.3±30.1 and 6.9±2.4 minutes, respectively • Left atrial dwell time was 29.6±15.3 minutes • 1 Primary SAE (cardiac perforation) and 2 AEs including vertigo (n=1; managed conservatively) and creatinine elevation (n=1; treated with IV saline). All AEs were resolved without sequelae
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360 Cardiac Catheter Status Expect to initiate a pivotal IDE for Paroxysmal AF in Q1 2026 ©2025 Pulse Biosciences, Inc. All rights reserved. Thyroid Therapy | Surgical Ablation | AF Ablation • Submitted FDA IDE application, including the pivotal study protocol • Treated over 150 patients in the multi-center feasibility study for the treatment of AF being conducted in Europe • Improved procedure times since last presented at the Heart Rhythm Society 2025 meeting 24
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©2025 Pulse Biosciences, Inc. All rights reserved. Summary Clinical evidence • Mounting and superior • Paradigm shifting care Expanding commercialization • Initiated nPulse Vybrance limited market release Portfolio of markets and pipeline of future indications • Multiple clinical or commercial programs activated IP +215 +100 • Pulse will own the Nanosecond PFA Space Novel nPulse Energy • Unique MOA • Patent Protected • Nonthermal U.S. clinical studies • 1 pivotal underway • 1 pivotal Q1 2026 • Measurable milestones Strong balance sheet • Cash balance of $95.2mm as of 9/30/2025 Target Market Values • +$6B U.S. Annual Addressable Mkt • Create, Expand, Upend 25
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KEEP RED LINE OFF SLIDE ©2025 Pulse Biosciences, Inc. All rights reserved. 26
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Citations 1) Data on file. Thyroidectomy WW Procedure Data provided by iData 2) https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8215427/ 3) Fine-Needle Aspiration of the Thyroid Gland https://www.ncbi.nlm.nih.gov/books/NBK285544/ 4) CMS - https://www.cms.gov/medicare-coverage-database/view/lcd.aspx?lcdid=38968&ver=4 5) CDC - https://seer.cancer.gov/statfacts/html/thyro.html 6) Global Cardiac Surgical Volume and Gaps: Trends, Targets, and Way Forward. Annals of Thoracic Surgery. 2023, ISSN 2772 -9931, https://doi.org/10.1016/j.atssr.2023.11.019. 7) Market size for 2023. 1% CAGR for Surgical Volume 8) Clarivate – US EP Market Report 9) Company filings, BofA Global Research. Revenue is BofA estimate 10) Neis et al. Circ Arrhythm Electrophysiol. 2024;17:e012854. DOI: 10.1161/CIRCEP.124.012854 11) Wyler von Ballmoos, Moritz C. et al.The Annals of Thoracic Surgery, Volume 117, Issue 2, 260 – 270 12) McCarthy, P. M. et al. Prevalence of atrial fibrillation before cardiac surgery and factors associated with concomitant ablat ion. J. Thorac. Cardiovasc. Surg. 159, 2245-2253.e15. 13) Burrage, P.S., Low, Y.H., Campbell, N.G. et al. New -Onset Atrial Fibrillation in Adult Patients After Cardiac Surgery. Curr Anesthesiol Rep 9, 174–193 (2019). https://doi.org/10.1007/s40140-019-00321- 4 14) Wong CX, Brown A, Tse HF, et al. Epidemiology of Atrial Fibrillation: The Australian and Asia -Pacific Perspective. Heart Lung Circ. 2017;26(9):807-879 15) Joglar et al J.A.C.C. V O L . 8 3 , N O . 1 , 2 0 2 42023 Guideline for the Diagnosis and Management of Atrial Fibrillation J A N U A R Y 2 / 9 , 2 0 2 4 : 1 0 9 – 2 7 9116 ( Linear Interpolation ) 16) Pulsed Field or Conventional Thermal Ablation for Paroxysmal Atrial Fibrillation. https://pubmed.ncbi.nlm.nih.gov/37634148/ 17) Lesion Morphometry of the Pentaspline Pulsed Field Ablation Catheter: Understanding Catheter Pose, Rotation, and Dosing. https://www.ahajournals.org/doi/epub/10.1161/CIRCEP.124.013208 18) Koruth et al. Circ Arrhythm Electrophysiol. 2024;17:e012854. DOI: 10.1161/CIRCEP.124.012854 ©2025 Pulse Biosciences, Inc. All rights reserved. 27