<?xml version="1.0"?>
<ownershipDocument>

    <schemaVersion>X0508</schemaVersion>

    <documentType>4</documentType>

    <periodOfReport>2025-06-01</periodOfReport>

    <issuer>
        <issuerCik>0001711933</issuerCik>
        <issuerName>Akoya Biosciences, Inc.</issuerName>
        <issuerTradingSymbol>AKYA</issuerTradingSymbol>
    </issuer>

    <reportingOwner>
        <reportingOwnerId>
            <rptOwnerCik>0002035474</rptOwnerCik>
            <rptOwnerName>BAMFORD PASCAL</rptOwnerName>
        </reportingOwnerId>
        <reportingOwnerAddress>
            <rptOwnerStreet1>C/O AKOYA BIOSCIENCES, INC.</rptOwnerStreet1>
            <rptOwnerStreet2>100 CAMPUS DRIVE, 6TH FLOOR</rptOwnerStreet2>
            <rptOwnerCity>MARLBOROUGH</rptOwnerCity>
            <rptOwnerState>MA</rptOwnerState>
            <rptOwnerZipCode>01752</rptOwnerZipCode>
            <rptOwnerStateDescription></rptOwnerStateDescription>
        </reportingOwnerAddress>
        <reportingOwnerRelationship>
            <isDirector>0</isDirector>
            <isOfficer>1</isOfficer>
            <isTenPercentOwner>0</isTenPercentOwner>
            <isOther>0</isOther>
            <officerTitle>Chief Clinical Officer</officerTitle>
        </reportingOwnerRelationship>
    </reportingOwner>

    <aff10b5One>0</aff10b5One>

    <nonDerivativeTable>
        <nonDerivativeTransaction>
            <securityTitle>
                <value>Common Stock</value>
            </securityTitle>
            <transactionDate>
                <value>2025-06-01</value>
            </transactionDate>
            <transactionCoding>
                <transactionFormType>4</transactionFormType>
                <transactionCode>F</transactionCode>
                <equitySwapInvolved>0</equitySwapInvolved>
            </transactionCoding>
            <transactionAmounts>
                <transactionShares>
                    <value>670</value>
                    <footnoteId id="F1"/>
                </transactionShares>
                <transactionPricePerShare>
                    <value>1.25</value>
                </transactionPricePerShare>
                <transactionAcquiredDisposedCode>
                    <value>D</value>
                </transactionAcquiredDisposedCode>
            </transactionAmounts>
            <postTransactionAmounts>
                <sharesOwnedFollowingTransaction>
                    <value>205738</value>
                </sharesOwnedFollowingTransaction>
            </postTransactionAmounts>
            <ownershipNature>
                <directOrIndirectOwnership>
                    <value>D</value>
                </directOrIndirectOwnership>
            </ownershipNature>
        </nonDerivativeTransaction>
    </nonDerivativeTable>

    <footnotes>
        <footnote id="F1">Represents shares withheld to cover taxes on a restricted stock unit award previously granted on May 18, 2023, which vests in four (4) annual equal installments beginning on June 1, 2024 and annually thereafter until fully vested.</footnote>
    </footnotes>

    <ownerSignature>
        <signatureName>/s/ Pascal Bamford by Brian McKelligon, Attorney-in-Fact</signatureName>
        <signatureDate>2025-06-02</signatureDate>
    </ownerSignature>
</ownershipDocument>
